Endemic pediatric fungal infections in India: clues to diagnosis.

This review is intended to familiarize readers with an emerging group of fungal infections that mostly manifest in immunocompetent individuals. This group was initially considered endemic to the tropics, but increasing worldwide prevalence has been reported. The organisms have been divided into dominant non-invasive forms and dominant invasive forms for ease of understanding. The non-invasive organisms include the group Entomophthoromycota, under which two genera Basidiobolus and Conidiobolus, have been identified as human pathogens. They present with plaques in the extremities and rhinofacial region, respectively. The invasive organisms are dematiaceous fungi (phaeohypomycosis), which includes Cladophialophora and Exophiala among others. They cause invasion of deep tissues, with the central nervous system being the most common target. The mycology, epidemiology, diagnosis, and treatment options have been summarized in brief. The clinical presentation, imaging manifestations, differentiation from other common infections and malignancies that show similar features have been detailed.

Octanoic acid kills Lucilia sericata (Diptera: Calliphoridae) by affecting two major defence systems: Cuticular free fatty acids and immunocompetent cells.

This work examines the insecticidal activity of octanoic acid (C8:0), a short-chain fatty acid detected in entomopathogenic fungus - Conidiobolus coronatus medium, against Lucilia sericata larvae and adults. The LD50 value was calculated as 3.04 ±â€¯0.26 µg/mg (3040 mg/kg) of insect body mass, which places the compound in category 5 of acute toxicity (slightly hazardous). The presented research also describes its probable mechanism, with a particular focus on changes in two main insect defense mechanisms: (1) the composition of the cuticle (GC-MS analysis) and (2) immunocompetent cells (microscopic analysis of cultured hemocytes). More precisely, octanoic acid application resulted in changes in cuticular free fatty acid (FFA) profiles in both adults and larvae; generally, treatment increased short-chain FFAs, and a decrease of middle- and long-chain FFAs. Both in vivo and in vitro applications of octanoic acid resulted in vacuolisation, disintegration, and destruction of nets formed by plasmatocytes. As the compound has also previously been found to be toxic against Galleria mellonella, it appears to have lethal potential against insects in both the Orders Diptera and Lepidoptera, indicating it may have strong entomopathogenic potential. It is worth noting that octanoic acid is approved as a food additive with well-documented insecticidal activity, and hence may be a valuable component in the design of new insecticides that are safe for both humans and the environment.

The effect of infection with the entomopathogenic fungus Conidiobolus coronatus (Entomopthorales) on eighteen cytokine-like proteins in Galleria mellonella (Lepidoptera) larvae.

Background: In response to the replace mammal research models with insects in preliminary immunological studies, interest has grown in invertebrate defense systems. The immunological response is regulated by cytokines; however, while their role in mammals is well understood, little is known of their function in insects. A suitable target for studies into insect immunology is Galleria mellonella (Lepidoptera), the wax moth: a common host for human fungal and bacterial pathogens. G. mellonella is also a perfect subject for studies into the presence of cytokine-like proteins. Specific objectives: The main goal of present research was detection in insect immunocompetent cells the 18 mammalian cytokines (IL-1α, IL-1ß, IL-2, IL-3, IL-6, IL-7, IL-8, IL-12, IL-13, IL-15, IL-17, IL-19, IFN-γ, TNF-α, TNF-ß, GM-CSF, M-CSF, G-CSF), which play important role in immunological response and indication how their level change after fungal infection. Methodology: The changes of cytokine-like proteins level were detected in hemocytes taken from G. mellonella larvae infected with entomopathogenic fungus, C. coronatus. The presence of cytokine-proteins was confirmed with using fluorescence microscopy (in cultured hemocytes) and flow cytometry (in freshly collected hemolymph). The ELISA test was used to detect changes in concentration of examined cytokine-like proteins. Results: Our findings indicated the presence of eighteen cytokine-like molecules in G. mellonella hemocytes during infection with C. coronatus. The hemocytes taken from infected larvae demonstrated higher fluorescence intensity for six cytokine-like proteins (GM-CSF, M-CSF, IL-3, IL-15, IL-1ß and IL-19) compared to untreated controls. ELISA test indicated significantly higher IL-3 and IL-15. M-CSF, IL-1α and IL-19 concentration in the hemolymph after fungal infection, and significantly lower TNF-ß and G-CSF. Conclusions: Our findings confirm that the selected cytokine-like molecules are present in insect hemocytes and that their concentrations change after fungal infection, which might suggest that they play a role in the anti-fungal immunological response.

Clinical characteristics of 5 adult patients with rhinofacial entomophthoromycosis in Thailand.

Five patients [mean age: 36 years (range: 22-65)] were diagnosed with rhinofacial entomophthoromycosis at our center during the study period. All patients presented with painless cheek and nasal dorsum swelling with nasal obstruction. All pathology reports confirmed rhinofacial entomophthoromycosis, which is characterized by the Splendore-Hoeppli phenomenon. Conidiobolus coronatus was identified from fungal culture in all patients. All patients were successfully treated with various antifungals.

The Entomopathogenic Fungus Conidiobolus coronatus Has Similar Effects on the Cuticular Free Fatty Acid Profile of Sensitive and Resistant Insects.

The mechanisms underlying the recognition of a susceptible host by a fungus and the role of cuticular compounds (CCs) in this process remain unclear; however, accumulated data suggest that this is influenced to a great degree by cuticular lipids. Two insect species differing in their sensitivity to fungal infection, viz. the highly sensitive Galleria mellonella Linnaeus (Lepidoptera: Pyralidae) and the resistant Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae), exhibited significant qualitative and quantitative changes in cuticular free fatty acid (FFA) profiles after exposure to Conidiobolus coronatus (Constantin) Batko (Entomopthorales). Despite being systematically distant, leading different lifestyles in different habitats, both insect species demonstrated similar changes in the same FFAs following exposure to the fungus (C12:0, C13:0, C14:0, C15:0, C16:1, C16:0, C18:1, C18:0), suggesting that these are involved in a contact-induced defense response. As it was not possible to distinguish the share of FFAs present in the conidia that were attached to the cuticle from the FFAs of the cuticle itself in the total number of extracted FFAs, further research is necessary.

Entomoftoromicosis por Conidiobolus coronatus en un paciente joven proveniente de área rural / Entomophthoromycosis due to Conidiobolus coronatus in a young patient from a rural area

La conidiobolomicosis es una micosis subcutánea causada por un hongo saprofito, Conidiobulus spp. perteneciente a la clase Zigomicetos, orden Entomoftorales, que habita en regiones tropicales. La manifestación clínica clásica es la deformidad progresiva de estructuras faciales y su diagnóstico se basa en cultivos de la zona afectada y el estudio histopatológico, siendo el "fenómeno de Splendore-Hoeppli" el hallazgo más característico. Dada su baja frecuencia de presentación, no existe consenso sobre la mejor opción y tiempo de tratamiento. Aquí presentamos un caso de entomoftoromicosis rinofacial causada por Conidiobolus coronatus en un paciente inmunocompetente de la región sur de Colombia.

Conidiobolomycosis is a subcutaneous mycosis caused by a saprophytic fungus, Conidiobulus, belonging to the class of Zygomycetes, an order of Entomophtorales that inhabits tropical regions. Its most frequent clinical manifestation is the progressive deformity of facial midline structures, and the diagnosis is based on cultures taken from the affected area and the histopathological study, being the "Splendore-Hoeppli phenomenon" the most characteristic finding. Due to its low frequency of presentation, there is no consensus about the best option and treatment time. We present a case of rhinofacial entomophthoromycosis caused by Conidiobolus coronatus in an immunocompetent patient from the southern region of Colombia.

Analysis of laccase-like enzymes secreted by fungi isolated from a cave in northern Spain.

Laccases belong to a family of multicopper enzymes able to oxidize a broad spectrum of organic compounds. Despite the well-known property of laccases to carry out bleaching and degradation of industrial dyes and polyphenolic compounds, their industrial use is often limited by the high cost, low efficiency, or instability of these enzymes. To look for new microorganisms which produce laccases that are potentially suitable for industrial applications, we have isolated several fungal strains from a cave in northern Spain. Their phenotypic analysis on agar plates supplemented with ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) disclosed two laccase-positive strains. Further genotyping revealed that they belonged to the Gliomastix murorum and Conidiobolus thromboides species. The secretion of G. murorum and C. thromboides laccase-like enzymes was then confirmed by zymography. Further identification of these polypeptides by mass-spectroscopy revealed the nature of the laccases and made it possible to predict their functional domains and other features. In addition, plate assays revealed that the laccases secreted by both G. murorum and C. thromboides were capable of degrading industrial dyes (Congo Red, Indigo, and Eriochrome Black T). Homology modeling and substrate docking predicted the putative structure of the currently uncrystallized G. murorum enzyme as well as its amino acid residues potentially involved in interactions with these dyes. In summary, new biochemical and structural insights into decolorization mediated by G. murorum laccase as well as identification of laccase-like oxidase in C. thromboides point to a promising future for these enzymes in biotechnology.

A rare case of condiobolomycosis due to Conidiobolus coronatus presenting with dysphagia.

Conidiobolomycosis is a neglected tropical fungal infection involving the head and neck region. Here we report the first case of atypical conidiobolomycosis presenting with dysphagia and significant weight loss from Odisha, India. It was diagnosed by histopathology and fungal culture and was suscessfully treated with saturated solution of potassium iodide.

Infection of Galleria mellonella (Lepidoptera) Larvae With the Entomopathogenic Fungus Conidiobolus coronatus (Entomophthorales) Induces Apoptosis of Hemocytes and Affects the Concentration of Eicosanoids in the Hemolymph.

Apoptosis and autophagy, the mechanisms of programmed cell death, play critical roles in physiological and pathological processes in both vertebrates and invertebrates. Apoptosis is also known to play an important role in the immune response, particularly in the context of entomopathogenic infection. Of the factors influencing the apoptotic process during infection, two of the lesser known groups are caspases and eicosanoids. The aim of this study was to determine whether infection by the entomopathogenic soil fungus Conidiobolus coronatus is associated with apoptosis and changes in caspase activity in the hemocytes of Galleria mellonella larvae, and to confirm whether fungal infection may affect eicosanoid levels in the host. Larvae were exposed for 24 h to fully grown and sporulating fungus. Hemolymph was collected either immediately after termination of exposure (F24 group) or 24 h later (F48 group). Apoptosis/necrosis tests were performed in hemocytes using fluorescence microscopy and flow cytometry, while ELISA tests were used to measure eicosanoid levels. Apoptosis and necrosis occurred to the same degree in F24, but necrosis predominated in F48. Fungal infection resulted in caspase activation, increased PGE1, PGE2, PGA1, PGF2α, and 8-iso-PGF2α levels and decreased TXB2 levels, but had no effect on TXA2 or 11-dehydro-TXB2 concentrations. In addition, infected larvae demonstrated significantly increased PLA2 activity, known to be involved in eicosanoid biosynthesis. Our findings indicate that fungal infection simultaneously induces apoptosis in insects and stimulates general caspase activity, and this may be correlated with changes in the concentrations of eicosanoids.

Case Report: Pulmonary Conidiobolomycosis in a Vietnamese Pot-Bellied Pig.

An adult castrated male Vietnamese pot-bellied pig had a 1-week history of acute dyspnea and lethargy. Minimal diagnostic testing was authorized by the owner, resulting in treatment with a third-generation cephalosporin and a non-steroidal anti-inflammatory drug. Partial improvement was observed after a week; however, the pig died 2 weeks after the initial onset of clinical signs. Macroscopically, ~90% of the left lung was effaced by large masses with a caseonecrotic center. Histologic examination revealed eosinophilic granulomas with myriad, intralesional, negatively staining hyphae highlighted by "sleeves" of hypereosinophilic material (Splendore-Hoeppli material). Infection with an oomycete or "zygomycete" (i.e., organisms of the order Entomophthorales or Mucorales) was initially considered. Pan-fungal PCR and sequencing performed on formalin-fixed, paraffin-embedded lung tissue identified Conidiobolus spp., consistent with a diagnosis of primary pulmonary conidiobolomycosis. There are only a few reports of infections with Conidiobolus spp. (and other members of the order Entomophthorales) in swine. Unlike humans and other animal species, conidiobolomycosis in pigs presents more commonly as a primary pulmonary disease rather than rhinofacial or nasopharyngeal disease.

Entomophthoramycosis: An unusual cause of facial disfigurement.

Entomophthoramycosis is a rare fungal infection of nose, paranasal sinuses and subcutaneous tissues found in tropical and subtropical region. From India very few cases have been reported. Here we report a case of Entomophthoramycosis due to Conidiobolus coronatus from the eastern India who presented with slowly growing rhinofacial swelling and right sided nasal obstruction due to intranasal mass. The case was diagnosed by typical histopathological findings of broad aseptate hyphae with surrounding eosinophilic granular material (Splendore Hoeppli phenomenon) on microscopy of nasal biopsy material and confirmed by PCR assay of DNA and sequencing from biopsy tissue. Treatment with saturated solution of potassium iodide and itraconazole was successful and clinical cure was attained in 8 months.

Branched-Chain Fatty Acids as Mediators of the Activation of Hepatic Peroxisome Proliferator-Activated Receptor Alpha by a Fungal Lipid Extract.

The study aimed to test the hypothesis that monomethyl branched-chain fatty acids (BCFAs) and a lipid extract of Conidiobolus heterosporus (CHLE), rich in monomethyl BCFAs, are able to activate the nuclear transcription factor peroxisome proliferator-activated receptor alpha (PPARalpha). Rat Fao cells were incubated with the monomethyl BCFAs 12-methyltridecanoic acid (MTriA), 12-methyltetradecanoic acid (MTA), isopalmitic acid (IPA) and 14-methylhexadecanoic acid (MHD), and the direct activation of PPARalpha was evaluated by reporter gene assay using a PPARalpha responsive reporter gene. Furthermore, Fao cells were incubated with different concentrations of the CHLE and PPARalpha activation was also evaluated by using the reporter gene assay, and by determining the mRNA concentrations of selected PPARalpha target genes by real-time RT-PCR. The reporter gene assay revealed that IPA and the CHLE, but not MTriA, MHD and MTA, activate the PPARalpha responsive reporter gene. CHLE dose-dependently increased mRNA concentrations of the PPARalpha target genes acyl-CoA oxidase (ACOX1), cytochrome P450 4A1 (CYP4A1), carnitine palmitoyltransferase 1A (CPT1A) and solute carrier family 22 (organic cation/carnitine transporter), member 5 (SLC22A5). In conclusion, the monomethyl BCFA IPA is a potent PPARalpha activator. CHLE activates PPARalpha-dependent gene expression in Fao cells, an effect that is possibly mediated by IPA.

Conidiobolomycosis, cryptococcosis, and aspergillosis in sheep and goats: a review.

We review herein infections by Conidiobolus spp., Cryptococcus spp., and Aspergillus spp. in sheep and goats. Conidiobolus spp. are common causes of rhinitis in sheep and are less frequent in goats, in which Conidiobolus spp. also cause skin lesions. Cryptococcus spp. cause rhinitis, meningitis, encephalitis, and pneumonia in goats, and are rarely observed in sheep. Aspergillus spp. may cause rhinitis in goats, and pneumonia and mastitis in sheep and goats. Gross and microscopic lesions caused by these 3 fungal infections may be similar to each other. The diagnosis of these diseases must be based on gross and microscopic lesions, coupled with detection of the agent by immunohistochemical, molecular, and/or culture-based methods.

Rhinofacial entomophthoramycosis case series, the unusual cause of facial swelling.

BACKGROUND: Rhinofacial entomophthoramycosis is a specific fungal infection of the skin and subcutaneous tissue. It is considered as a rare and neglected disease in tropical and subtropical areas. We would like to present our cases to aid other physicians in the improved recognition of typical cases. MATERIALS AND METHODS: A retrospective review was performed on patients with the diagnosis of Conidiobolomycosis or Entomophthoramycosis in Chiang Mai University Hospital, Thailand, from January 2009 to May 2019. There were seven cases with a definite pathologic report or culture in this review. RESULTS: All seven patients were men and were referred to the university hospital for diagnosis. The mean age was 53 ± 15.7, ranging from 27 to 71 years. Most of the patients (85.7%) presented first with nasal or rhinofacial swelling and nasal obstruction. The definite diagnosis came from clinical presentation and investigation with a tissue biopsy, culture and communication among physicians. Patients responded well with a combination of medical treatment, including potassium iodide (KI), co-trimoxazole, or itraconazole. CONCLUSION: Rhinofacial entomophthoromycosis or Conidiobolomycosis typically can be diagnosed under a suspicious clinical presentation. The obvious clinical response can be seen within several weeks after medication.

Conidiobolus, a hitherto unidentified pathogen in microbial keratitis.

Fungal infections are a significantly increasing cause of ocular and systemic morbidity; the vast majority of cases being ascribed to a handful of species. Fungal keratitis, unlike systemic infections, usually occur in immunocompetent individuals. Rarely, systemic infections can be associated with ocular involvement (e.g., Candida, Mucor, Pythium), or a fungus that predominantly causes systemic disease can affect the eye. One such fungus is Conidiobolus which is known to cause muco-cutaneous infections. We report the identification and successful treatment of a case of Conidiobolus corneal ulcer in an immunocompetent individual, who had no co-existing muco-cutaneous disease. Identification of this particular fungus and awareness of its potential to cause systemic disease is especially relevant, given its potential for chronic indolent infection of the subcutaneous tissues. To the best of our knowledge, this is the first reported case of a Conidiobolus corneal ulcer.

Genome-wide study of saprotrophy-related genes in the basal fungus Conidiobolus heterosporus.

Conidiobolus spp. are important saprophytic basal fungi. However, to date, no genomic-level data for decaying plant materials in the genus Conidiobolus has been reported. Here, we report that the 33.4-Mb genome of Conidiobolus heterosporus encodes 10,857 predicted genes. Conidiobolus heterosporus harbors 394 CAZyme-encoding genes belonging to 4 major modules but does not encode a polysaccharide lyase (PL). Many carbohydrate esterases (CEs) belonging to the family CE12 play crucial roles as pectin acetylesterases, and 14 genes were upregulated in the IM (fungus grown on inducing medium) among 17 expressed CE12 family genes. In addition, most of the genes in the GH132 CAZyme family showed a greater than 5-fold increase in expression in the IM compared with that in the wild type. Furthermore, 122 P450-encoding genes grouped into 11 families were detected in the fungal genome, most of which belonged to the CYP547 family (36 genes) followed by CYP548 (27 genes) and CYP5856 (25 genes). Interestingly, members of the families CYP5014 and CYP5136 were identified, the first time such enzymes have been described in a fungus. Our findings provide new insights into the genomics and genomic features of the saprophytic basal fungus C. heterosporus.Key Points• Genome of the saprobiotic basal fungus C. heterosporus was sequenced and analyzed.• 394 CAZymes but no PL family genes were found and expression levels were determined.• CE12 and GH132 proteins may play roles in the pectin and plant material degradation.• A large number of P450s but few P450 families existed in the fungus.

Nasofacial rhinoentomophthoramycosis: A report of two cases from Central India.

Rhinoentomophthoromycosis due to Conidiobolus coronatus is a rare, chronic, granulomatous disease, occurring mainly in tropical countries including India. We report two cases of rhinoentomophthoromycosis in an 18-year-old female and a farmer of 35 years residents of Chhattisgarh shifted from Madhya Pradesh and Orrisa. It was diagnosed by microscopy and isolation C. coronatus in culture. The patient presented with a swollen nose with obstruction that had progressed slowly over 1 year. His nasal swelling was bilateral, diffuse, mildly tender, erythematous, nonpitting, with mucosal crusting and hypertrophy of inferior turbinates but no regional lymphadenopathy. Culture of tissue from the nasal biopsy on sabouraud dextrose agar yielded multiple colonies of a mold with satellite smaller colonies at periphery. The isolate demonstrated the macroscopic and microscopic morphologic characteristics of C. coronatus. The patients were earlier treated with itraconazole or its combination with potassium iodide and the patients were treated successfully with amphotericin B.

Conidiobolomycosis: An Unusual Fungal Disease-Our Experience.

Conidiobolomycosis is a rare mycotic disease caused by Conidiobolus coronatus. Very few cases have been reported in English literature. Often it is clinically misdiagnosed as soft tissue tumour. A prospective case study was done from 2006 to 2015 in a tertiary care hospital of West Bengal, India. The objectives of our study were to describe the epidemiological and clinical features and treatment of Conidiobolomycosis to prevent disfigurement. Patients clinically suspected to be suffering from Conidiobolomycosis were subjected to biopsy followed by histopathological and mycological examinations. Then they were treated with oral saturated solution of potassium iodide along with other drugs. Total six cases were histopathologically proved to be suffering from Conidiobolomycosis. Fungus was isolated and identified in one case. Complete resolution was seen in five patients. Conidiobolomycosis should be brought into mind as differential diagnosis of subcutaneous swelling in the rhinofacial region.

Chronic invasive rhinosinusitis by Conidiobolus coronatus, an emerging microorganism.

Fungi from the Conidiobolus genus have been implicated in the development of chronic invasive fungal rhinosinusitis, mainly in tropical countries. The mycosis associated to these fungi may cause irreversible facial deformities and is potentially fatal. The authors present the first case of a chronic invasive fungal rhinosinusitis due to Conidiobolus coronatus diagnosed in a 66-year-old Caucasian male patient, living in Portugal without any travels abroad and complaining of progressive refractory nasal obstruction, facial pain and anosmia. Upon the culture of samples collected during sinus endoscopic surgery, colonies that presented a macroscopic aspect remembering wax were detected. The microscopic evaluation allowed the observation of simple conidiophores forming spherical conidia, and of conidiophores that presented hair-like appendages. Together, these characteristics allowed the identification of the fungi as a Conidiobolus, which was confirmed upon the DNA sequencing. The authors emphasised the role of this fungi as an emergent microorganism as well as the difficulties associated to the diagnosis and treatment.

Characterization and phylogenetic analysis of the complete mitochondrial genome of Conidiobolus sp. (Entomophthorales: Ancylistaceae).

In the present study, we presented the complete mitochondrial genome of an entomophthoroid fungus Conidiobolus sp. The mitogenome of Conidiobolus sp. has a total length of 26,612 bp, with the base composition as follows: A (44.22%), T (27.10%), C (10.99%) and G (17.68%). The mitogenome contains 19 protein-coding genes, 2 ribosomal RNA genes (rRNA), and 23 transfer RNA (tRNA) genes. The taxonomic status of the Conidiobolus sp. mitogenome exhibited a close relationship with the mitogenome of Conidiobolus heterosporus.