Fungal Stains Research Articles

Development of a Rapid Diagnostic Method for Sporotrichosis.

Sporotrichosis, a prevalent deep fungal infection in clinical settings, currently lacks rapid and accurate diagnostic methodologies. This study explores a novel rapid diagnosis method for sporotrichosis by combining FTA cards and nested PCR with fungal fluorescence staining. The study involved skin lesion tissues from 26 patients diagnosed with sporotrichosis (Experimental Group). The Positive Control Group consisted of fungal suspensions from clinical strains of Sporothrix, while the Negative Control Group included fungal solutions of other fungi, namely Trichophyton rubrum, Trichophyton mentagrophyte, and Candida albicans. DNA was extracted from the slurry of skin lesions in the Experimental Group and from fungal suspensions in the Control Group using FTA cards, followed by nested PCR amplification. Subsequently, nested PCR amplification was performed. Histopathological examinations, including HE and fluorescence staining, were conducted on paraffin sections prepared from skin lesion tissues in the Experimental Group. Among the 26 clinical skin lesion tissues in the Experimental Group, 8 cases showed a specific positive band upon nested PCR amplification, resulting in a positive rate of 30.8%. In the Control Group, the fungal solution of the clinical strain of Sporothrix showed a specific positive band upon nested PCR amplification, while all other fungi Negative Control Group tested negative. Histopathological examination revealed granulomatous inflammatory changes in most samples after HE staining. Fluorescence staining detected spores in 17 cases, resulting in a detection rate of 65.4% (17/26). The combination of FTA cards with nested PCR method proved to be simple and rapid but demonstrated a relatively low positive rate. Fungal fluorescence staining significantly improved the sensitivity of detecting sporotrichosis in histopathological examinations, thereby improving the speed and efficiency of diagnosis.

Expediting the bioactivity of zinc sulfide nanoparticles with copper oxide as a nanocomposite

The regulatory role of zinc in bone formation extends to the activation of proteins associated with bone homeostasis. Furthermore, copper is well known for its antibacterial properties. This dual function underscores the significance of zinc and copper in maintaining a balance of bone structure and function. In light of the aforementioned, zinc sulphide/copper oxide nanocomposites were created in this instance using a straightforward coprecipitation technique. Copper oxide was used as a nanocomposite to improve the structural, morphological, and biological performance of zinc sulphide nanoparticles. The X-ray diffraction pattern confirmed a transformation in the crystal structure from cubic to rhombohedral, along with increase in intensity. Fourier transforms infrared analysis indicated the presence of functional groups. Scanning electron microscopy images demonstrated a morphological shift from non-uniform to distinct spherical nanoparticles, impacting the enhancement of material properties. The pathogenic activity of the zinc sulphide/copper oxide nanocomposites was tested against nine bacterial strains. In antimicrobial testing, zinc sulphide/copper oxide nanocomposites showed promising results, particularly against Klebsiella pneumoniae (zone of inhibition: 14 mm at 100 µg/mL compared to 7 mm by standard) and Escherichia coli (zone of inhibition: 11 mm at 100 µg/mL compared to 10 mm by standard) after 24 h with zone of inhibition matching or exceeding that of the standard (chloramphenicol). Zinc sulphide nanoparticles and zinc sulphide/copper oxide nanocomposites were evaluated for their antifungal activity against fungal stains from Trichophyton rubrum, Aspergillus niger, and Aspergillus flavus. After a 24-h period, it was discovered that zinc sulphide/copper oxide nanocomposites were effective against Aspergillus flavus (zone of inhibition: 19.4 mm at 100 µg/mL compared to 6.3 mm by standard) at all concentrations (25–100 mg/mL), with zones of inhibition identical to or greater than those of the standard (fluconazole). Certainly, based on these results, zinc sulphide/copper oxide nanocomposites could be promising materials for drug delivery.Clinical trial registration: Not applicable.

Chronic Invasive Fungal Sinusitis Mimicking Malignancy Post-Radiotherapy: A Case Report.

Invasive fungal sinusitis is a life-threatening form of fungal rhinosinusitis. Due to the aggressive clinical presentation and radiological appearance, there is diagnostic difficulty in differentiating invasive fungal sinusitis from a malignant process. This is even more challenging in oncological patients who have undergone previous head and neck radiotherapy, due to possibility of a recurrence of primary malignancy and radiation-induced neoplasms. We report a rare case of invasive fungal sinusitis mimicking a malignancy in a post-radiotherapy patient.Our patient was a 68-year-old male, 25-years post-radiotherapy for nasopharyngeal carcinoma. He presented with a 3-month history of purulent sputum and right facial paraesthesia. Magnetic resonance imaging showed an irregular destructive enhancing mass of the greater wing of right sphenoid and pterygoid bone with extensive extension into nearby structures. In view of extensive local and bony invasion, and a history of radiotherapy, initial suspicions were that of primary malignancy, specifically radiation-induced sarcoma, and recurrence of nasopharyngeal carcinoma. He underwent transpterygoid biopsy of the lesion, and histopathology demonstrated Aspergillus species, with no malignancy identified.Our report highlights the diagnostic difficulties in the post-radiotherapy cancer patient presenting with symptoms suggestive of aggressive sino-nasal disease. Invasive fungal sinusitis closely mimics the clinical and radiological findings of several neoplastic processes. We discuss the clinical and radiological characteristics of pathologies that may mimic invasive fungal sinusitis. Histological examination remains the gold standard for diagnosis, and early fungal staining is crucial. Furthermore, one should not presume the initial histopathological diagnosis to be confirmatory of isolated fungal disease. Repeat radiological investigations for disease resolution and histopathologic re-evaluation if required should be performed, keeping in mind possibility of coexisting malignancy.

Decolorization of Textile Dyes by Extracellular Enzymes Produced from Trametes sanguinea and Perenniporia taephropora Immobilized on Natural Media

The color of textile wastewater is still a main problem in wastewater treatment by biological processes. The colored effluents from textile factories usually exceed effluent standards. Therefore, various innovations were developed to treat textile wastewater for decolorization in the effluents. This research aims to decolorize textile wastewater by immobilizing white rot fungi degradation. At first, the 11 fungal stains were tested to find the decolorized efficiency then the high decolorized efficiency fungal stains were immobilized on four material media, namely water hyacinth stalks, coconut husk, corn cob, and loofah. After that, the immobilized fungi were cultivated in the culture media at 30, 60, and 120 C/N ratios, respectively. The results showed that Trametes sanguinea and Perenniporia tephropora were two stains with a high decolorized efficiency of 68.8% and 67.5% respectively, and the decolorized efficiency was increased when immobilized on loofahs and fed with 120 C/N ratio medium. In a comparison of two fungal stains, P. tephropora was found more suitable for the decolorization of textile wastewater than T. sanguinea because T. sanguinea could produce red-orange pigments that induced the colored enhancement in wastewater over time. Finally, immobilized P. tephropora was cultivated in a 120 C/N ratio medium within a 10 L continuous stirred tank reactor (8 L working volume) to investigate the decolorized efficiency, enzymatic activity, and repeated batch. It was found that three repeated cycles were carried out by reusing the immobilized P. tephropora and the highest decolorized efficiency was 63.4%. The enzymatic activity of laccase, manganese peroxidase, and lignin peroxidase was 15.5 U/L, 85.9 U/L, and 0 U/L, respectively

Benzothiazole-based Schiff base for sensing Ca2+ ions: Synthesis, DFT studies, toxicity evaluation in zebrafish embryo and in silico analysis of MMP-9 inhibition

A new heterocyclic Schiff base, 2-(2-benzo(d)thiazol-2yl)hydrazono)methyl)phenol (BTAZ) synthesized from 2-hydrazinobenzothaizole and salicyclaldehyde was characterized by spectral, mass and single crystal XRD data and subjected to sense Ca2+ ions in methanol–water mixture (1:1, v/v) using UV–visible absorption and fluorescence emission spectral studies. The probe BTAZ showed specific detection of Ca2+ ions in a solution containing various metal ions with a detection limit as low as 5.1 × 10−6 M via photo-induced electron transfer (PET) pathway. The binding stoichiometry between BTAZ and Ca2+ was determined to be 2:1 by Job’s plot method. The stoichiometry and optical properties of both BTAZ and BTAZ-Ca2+ complex showed good correlation with theoretical results based on DFT and TD-DFT calculations. The pharmacological evaluation of the probe BTAZ against bacterial and fungal stains also demonstrated its notable efficacy to control their growth in vitro along with anti-oxidant capabilities. Further, toxicity assay of the probe BTAZ against zebrafish embryo did demonstrate that it is non-toxic to the test model in vivo. Ability of the probe BTAZ to inhibit matrix metalloproteinase-9 (MMP-9) enzyme was analyzed in detail using in silico experiments.

Clinico-Mycological Study of Dermatophytosis among Diabetic and Non-Diabetic Patients in a Tertiary Level Hospital: A Comparative Study

Abstract Background: Dermatophytosis is a major public health concern in India, especially in recent years, with an alarmingly rising trend, particularly in relation to recurrent and chronic infection. The number of studies examining the relationship between an individual’s glycemic status and the development of dermatophytosis, and a comparison of the evolving species trend between diabetic and non-diabetic patients infected with dermatophytes, is limited. Aims and Objectives: To study and compare the clinical and mycological aspects of dermatophytosis among diabetic and non-diabetic patients and to compare the culture isolates in both groups. Materials and Methods: The study included 378 patients of clinically suspected dermatophytosis, divided into two groups of 189 known diabetics and non-diabetics each. We subsequently analysed and compared the demographics, clinical data, potassium hydroxide (KOH) mount, calcofluor white (CW) staining and fungal culture results of all patients in both groups. Results: Among the 378 patients, diabetic patients had a significantly greater extent of involvement and higher rates of recurrence when compared to non-diabetics. The overall positivity rate was higher in CW staining (77.8%) as compared to KOH (57.7%) in both non-diabetics and diabetics. Trichophyton mentagrophytes was the most common isolate (47.6%) in both diabetic (55.0%) and non-diabetic (61.4%) patients, followed by Trichophyton rubrum (31.8% and 29.6%, respectively). The positivity percentages of fungal culture, KOH and CW staining increased to 100%, 77.9% and 95.7%, respectively, when scrapings were taken from ≥2 sites. Conclusion: The pattern of dermatophytosis is comparatively more chronic and severe in diabetics compared to non-diabetics. T. mentagrophytes was the most common culture isolate in both groups. CW staining can potentially be used as the initial method of choice for the diagnosis of dermatophytosis as it has significantly outperformed the conventional KOH mount. Scrapings can regularly be taken from ≥2 sites to avoid false negative results.

Ophiostoma haidaense, sp. nov., a new member of the O. piceae species complex associated with yellow-cedar, Callitropsis nootkatensis

ABSTRACT Ophiostoma haidanensis is described as a new species of the Ophiostoma piceae complex isolated from yellow-cedar (Callitropsis nootkatensis (D. Don) Oerst. ex D.P. Little) sapwood in the Haida Gwaii island archipelago and the North Coast of British Columbia, Canada. The fungus is characterized by the production of a typical sporothrix-like asexual morph but is distinguished morphologically from other members of the O. piceae species complex by its large, multiseptate primary conidia. Phylogenetic analysis of DNA sequences from the nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and the β-tubulin (BTUB) and translation elongation factor 1-α (TEF1) genes supports the inclusion of O. haidensis as a distinct member within the O. piceae complex. To our knowledge, this is the first report of a blue stain fungus infecting yellow-cedar, an ecologically, culturally, and economically important conifer naturally distributed along the coastal forests of the Pacific Northwest in North America.

Isolation and Diagnosis of Bacteria and Fungi from Some Areas of Tikrit and Some Villages

The purpose of the study was to evaluate the amount of air pollution caused by bacteria and fungi in specific areas of Tikrit, including the Al-Alam area, Al-Bu Ajil, Al-Buhyazaa, and the village of Al-Karaat. The study was conducted in November and January of 2021–2022, using petri dishes filled with nutrient agar (potato dextrose agar, or PDA) distributed throughout the study areas. The petri dish was left open for fifteen minutes before being closed and incubated with an incubator. After that, the dish was closed and the species was allowed to grow on the media, including isolates and differential media like Macconkey agar and Eosin Methylene Blue (EMB). For microscopical diagnosis, gram stain was applied to the bacterial samples, and fungal stain Lactophenol was used for fungus isolates. The findings of the current study showed that the proportion of Penicillium sp colonies colonized agricultural areas was twice as high, particularly in the Albu-Ajil area, where the dish G2.1 had more than 96 colonies, which was twice or three times more than the rest of the petri dishes in the study area for remote areas. This is because the area had ideal growth conditions in terms of temperature, humidity, and other factorsThe analysis revealed that the majority of bacteria were negative, particularly those belonging to the genus Klebsiella sp. which showed that it could grow on a Macconkey agar dish in Tikrit, the village of Al-Karaat, and Al-Alam. This suggests that it can grow in a variety of environments. However, in the Albu Hayaza area, no bacterial growth was found on the Macconkey agar dish, which was designated with the symbol 3.1 H. This could be because the environmental conditions at the time were insufficient for bacterial growth.

Isolation and Diagnosis of Bacteria and Fungi from Some Areas of Tikrit and Some Villages

The purpose of the study was to evaluate the amount of air pollution caused by bacteria and fungi in specific areas of Tikrit, including the Al-Alam area, Al-Bu Ajil, Al-Buhyazaa, and the village of Al-Karaat. The study was conducted in November and January of 2021–2022, using petri dishes filled with nutrient agar (potato dextrose agar, or PDA) distributed throughout the study areas. The petri dish was left open for fifteen minutes before being closed and incubated with an incubator. After that, the dish was closed and the species was allowed to grow on the media, including isolates and differential media like Macconkey agar and Eosin Methylene Blue (EMB). For microscopical diagnosis, gram stain was applied to the bacterial samples, and fungal stain Lactophenol was used for fungus isolates. The findings of the current study showed that the proportion of Penicillium sp colonies colonized agricultural areas was twice as high, particularly in the Albu-Ajil area, where the dish G2.1 had more than 96 colonies, which was twice or three times more than the rest of the petri dishes in the study area for remote areas. This is because the area had ideal growth conditions in terms of temperature, humidity, and other factorsThe analysis revealed that the majority of bacteria were negative, particularly those belonging to the genus Klebsiella sp. which showed that it could grow on a Macconkey agar dish in Tikrit, the village of Al-Karaat, and Al-Alam. This suggests that it can grow in a variety of environments. However, in the Albu Hayaza area, no bacterial growth was found on the Macconkey agar dish, which was designated with the symbol 3.1 H. This could be because the environmental conditions at the time were insufficient for bacterial growth.

Bioremediation of petroleum refinery wastewater by fungal stains isolated from the fishing harbour of Bizerte (Mediterranean Sea).

The study was conducted in order to explore the potential of fungi isolated from surface and bottom seawater collected from the fishing harbour of Bizerte on the bioremediation of industrial effluent (IE) contaminated by petroleum hydrocarbon. Among the 128 fungal isolates, 11 were isolated from surface seawater and 7 from bottom seawater, representing 18 taxa in total. The gas chromatography mass spectrometry (GC-MS) was used for the determination of hydrocarbon compounds in IE. An initial screening of fungal growth using six concentrations ranged between 20 and 70% (v/v) IE has allowed the identification of the optimal concentration for fungal growth as well as selection of species able to tolerate high amounts of hydrocarbon. Colorimetric test employing 2,6-dichlorophenol indophenol and gravimetric method was applied for the assessment of fungal growth using 20% EI. By checking the phylogenetic affiliation of the high-performing stains as identified using ITSr DNA sequence, a dominance of Ascomycetes was detected. Indeed, Aspergillus terreus and Penicillium expansum may degrade 82.07 and 81.76% of residual total petroleum hydrocarbon (TPH), respectively. Both species were collected from surface seawater. While, Aspergillus niger, Colletotrichum sp and Fusarium annulatum displayed comparable degradation rates 40.43%, 41.3%, and 42.03%, respectively. The lowest rate of degradation 33.62% was detected in Emericellopsis phycophila. All those species were isolated from bottom seawater, excepting A. niger isolated from surface water. This work highlighted the importance of exploring the potential of fungi isolated from the natural environment on the bioremediation of industrial effluent. Our results promoted the investigation of the potential of the high-performing isolates A. terreus and P. expansum on the bioremediation of IE at pilot-scale and then in situ.

Leach-resistant modified bamboo with fire-retardant and anti-mildew properties via in situ synthesis

To ensure the long-term fire-retardant (FR) and anti-mildew (AM) of bamboo materials when employed in outdoor settings susceptible to rainfall exposure, high durability is essential. In this study, we achieved water-leaching resistance of FR and AM bamboo through the in situ synthesis of phosphotungstic acid (PW) and silver nitrate (AG) via vacuum-pressure impregnation. The successful synthesis of the nanospheres within the bamboo (referred to as PWAG-B) was confirmed by TEM, FTIR, and XPS. These nanospheres remained intact within the bamboo cell wall after being subjected to a leaching test, with only a negligible mass loss of 0.88% and virtually no traces of Ag and W in the leachate for PWAG-B1. Furthermore, the treated bamboo exhibited superior AM properties, with complete resistance against two types of mildew, namely Botryodiplodia theobromae and Alternaria alternate, as well as stain fungi including Aspergillus niger and Penicillum citrinum. Notably, the treated bamboo displayed reduced thermal and smoke parameters in comparison to untreated bamboo, along with a delayed ignition time. This long-term AM and FR bamboo has the potential for the development of eco-friendly outdoor building materials.

Molecular Identification and Phytochemical Analysis and Bioactivity Assessment of Catharanthus roseusLeaf Extract: Exploring Antioxidant Potential and Antimicrobial Activities.

Plants have long been at the main focus of the medical industry's attention due to their extensive list of biological and therapeutic properties and ethnobotanical applications. Catharanthus roseus, sometimes referred to as Nithyakalyani in Tamil, is an Apocynaceae family member used in traditional Indian medicine. It also examines the plant's potential antimicrobial and antioxidant activities as well as its preliminary phytochemical makeup. Leaf material from C. roseus was analyzed and found to include a variety of phytochemicals including alkaloids, terpenoids, flavonoids, tannins, phenols, saponins, glycosides, quinones, and steroids. Four of the seven secondary metabolic products discovered in C. roseus leaves showed bioactive principles: 3-methylmannoside, squalene, pentatriacontane, and 2,4,4-trimethyl-3-hydroxymethyl-5a-(3-methyl-but-2-enyl)-cyclohexene. Catharanthus roseus is rich in the anticancer compounds vinblastine and vincristine. Whole DNA was isolated from fresh leaves, then amplified, sequenced, and aligned to find prospective DNA barcode candidates. One DNA marker revealed the restricted genetic relationship among C. roseus based on genetic distance and phylogenetic analysis. The antioxidant activity of the plant extract was evaluated using the DPPH, ABTS, phosphomolybdenum, FRAP, and superoxide radical scavenging activity assays, while the antibacterial potential was evaluated using the agar well diffusion assay. The ethanol extract of C. roseus was found to have the highest reducing power. In addition, a 4- to 21-mm-wide zone of inhibition was seen when the C. roseus extract was tested against bacterial and fungal stains. In conclusion, C. roseus has the most promise as an antibacterial and antioxidant agent.

Promjena boje površina drva izloženih vremenskim utjecajima prije i nakon tretmana željezovim (II) sulfatom

Outdoor wood is exposed to various factors that can be described as weathering and cause the wood to grey. The ageing processes can vary greatly depending on the exposure. Parts of wood that are not exposed to external factors, e.g. under the overhanging, are less exposed, and the weathering process is therefore slower. This can be accelerated by solutions based on iron ions. In this way, the wood greys quickly and evenly. However, after iron treatment, the colour also depends on the previous exposure of wood to weathering. In our study, the colour change was observed as a function of weathering time. Before treatment, the samples were exposed to outdoor weathering for different periods of time and then treated with a 5 % solution of pure iron (II) sulphate and commercial iron (II) sulphate. It was determined that the pre-weathering time affected the final colour change, as the samples exposed for five weeks before treatment have comparable colour to naturally weathered wood. At the beginning of exposure, iron (II) sulphate limits mould growth, but after two months, staining fungi develop on the treated samples as well. The growth of blue stain fungi on the treated samples did not significantly affect the colour and visual appearance of the wood treated with iron-based solutions.

Efficient crop straws biotreatment using the fungus Cerrena Unicolor GC.u01

Lignin is main composition of agricultural biomass which can be decomposed through enzymatic hydrolysis by fungi. However, there are still needs to identify more efficient and effective fungal stain for biomass valorization. In this study, lignin degrading fungi from birch forest were screened for sustainable degradation of waste agricultural straws. The most effective strain was identified as Cerrena unicolor GC.u01 using 18 S rDNA gene-sequencing technology. Three different crop straws (corn stalk, rice and wheat straws) were used for the biotreatment studies. The activities of lignin degrading enzymes, laccase (Lac), cellulase and xylanase, secreted by C. unicolor were also determined. Scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FTIR) and thermal gravimetric analyzer (TGA) were further used to monitor the effects of the biotreatment process. The results showed that C. unicolor degraded 34.3% rice straw lignin, a percentage which was higher than other isolated strains after 15 d straw liquid fermentation. The highest Lac activity (8.396 U•mL− 1) was observed with corn stalk on the 7 d. Cellulase and xylanase activities, in the same biomass, were higher than those of wheat and rice straws after 15 d. Furthermore, SEM, FTIR and TGA analyses showed that C. unicolor pretreatment process had significant effects on corn stalk, rice and wheat straws’ structures. The newly isolated stain of C. unicolor demonstrated high lignin degradation potential that can provide effective, ecofriendly means of valorizing biomass to industrial useable raw-material.

A diverse spectrum of mycoses histologically diagnosed in Ghana: Insights from a 10-year retrospective study.

In Ghana, most laboratory diagnoses of severe mycoses are based on histopathology findings due to inadequate availability of serology, culture, and molecular tests. The aim of this study was to evaluate the spectrum of mycoses diagnosed in Ghana. We retrospectively reviewed reports from 2012 to 2021 from three major pathology laboratories in Ghana to identify reports indicating the presence of fungal elements and diagnosis of a mycosis, then extracted demographic, clinical history, site of infection, stain(s), used and diagnosed mycosis details. Over the 10-year period, 107 cases were found. No apparent increasing and decreasing trend in the number of cases per year or in a period was observed. The age range of affected patients was from 4 to 86 years. Special stains for fungi were only used in 22 of 107 (20.6%) of cases. The most frequently affected site was the sino-nasal area (34%). Mycosis type was determined for 58 (54.2%) cases, comprising aspergillosis (21), candidiasis (14), dermatophytosis (6), mucormycosis (3), two cases each of chromoblastomycosis, histoplasmosis, eumycetoma, entomophthoromycosis, sporotrichosis, and Malassezia infection and a single case each of cryptococcosis and deep onychomycosis. Of the 53 (49.5%) cases with presumptive diagnosis data, only seven (13.2%) had a pre-biopsy suspicion of mycosis. There is a wide spectrum of mycoses in Ghana, including endemic mycoses not previously reported. Improving the use of special fungal stains could increase yield and mycoses identification. Laboratory diagnostic capacity needs enhancement to complement histopathology investigations with serology, culture, and molecular methods.

Efficacy and Rapidity of Potassium Hydroxide Mount and Modified Chicago Sky Blue 6B Stain with Potassium Hydroxide in Fungal Keratitis Detection.

To compare the efficacy and rapidity of direct microscopic detection of fungal elements from corneal ulcers between 10% potassium hydroxide (KOH) and 1% Chicago Sky Blue 6B (CSB) in 10% KOH (CSB-KOH). Thirty patients with clinically suspected fungal keratitis were recruited. Participants with impending corneal perforation were excluded. Two slides were smeared with corneal ulcer scrapings from the ulcer's edge and base for comparison of fungal staining solutions. One slide was infused with KOH, and the other slide was filled with CSB-KOH. Additional scraping was collected for inoculation on Sabouraud dextrose agar for fungal culture. The sensitivity, specificity and rapidity of both stainings were analyzed. The sensitivity of fungal culture, KOH, and CSB-KOH were 43.75% (95% confidence interval [CI], 19.75%-70.12%), 62.50% (95% CI, 35.43%-84.80%), and 87.50% (95% CI, 61.65%-98.45%), respectively. The specificity were 100% (95% CI, 69.15%-100%) of both stainings and fungal culture which analyzed from 16 fungal keratitis cases by laboratory and clinical diagnosis. Mean CSB-KOH examination time was quicker than KOH with the mean time difference of 5.6 minutes (95% CI, 3.22-7.98 minutes) and p-value < 0.001. CSB-KOH was more effective and faster than KOH in detecting fungal elements from corneal ulcers. Therefore, CSB-KOH may be beneficial in diagnosing fungal keratitis and preventing blindness. Moreover, to the best of our knowledge, this is the first use of CSB stain in fungal keratitis detection.

Effects of Different Levels of Physical Damage Combined with Fungal Induction on Agarwood Formation

As wild Aquilaria sinensis resources are exhausted and protected, China has established a huge number of plantations of Aquilaria trees and developed artificial induction techniques. However, the current output and quality of artificial induction technology have not yet met the expected results. It has been found that high-oil-containing agarwood may contain particular fungal stains associated with agarwood production. To enhance the quality of agarwood, we recovered and characterized three highly active fungi from high-oil-containing agarwood and inoculated them onto A. sinensis trees using two traditional physical methods. The results showed that fungi extracted from high-oil-containing agarwood can effectively increase the yield and quality of agarwood. During the agarwood formation process, parenchyma cells, xylem rays, and axial parenchyma cells in the xylem gradually undergo apoptosis, thereby promoting the expansion of the color range of agarwood. Nine months after the treatment, the alcohol-soluble extract content in agarwood reached the standard specified in the Chinese Pharmacopoeia (10%), and the proportions of sesquiterpenes and chromones in each treatment were 55.82%, 58.31%, 62.65%, 70.97%, and 13.71%, respectively. These results indicate that fungal induction has a positive impact on the quality of agarwood. In addition, compared to drilling and fungus combined induction, “burning holes and fungi” combined induction demonstrates better results and can further improve the yield and quality of agarwood.

Expert consensus on diagnosis and treatment of severe COVID-19 associated pulmonary aspergillosis and mucormycosis

The incidence and mortality of COVID-19 associated pulmonary aspergillosis (CAPA) are high in critically ill patients. Although COVID-19 associated mucormycosis (CAPM) is relatively rare, its severity and often a delayed diagnosis or misdiagnosis lead to its high mortality. The diagnosis and treatment of CAPA and CAPM in critically ill patients are challenging. Early diagnosis and a standardized therapy are the two most important factors for a good outcome. Therefore, a working group of experts from Chinese Thoracic Society and Chinese Association of Chest Physicians Critical Care Group was organized to develop this consensus based on the current medical evidence and clinical practice, in order to improve the ability of clinical treatment for critically ill patients with CAPA and CAPM. The working group drafted a preliminary text based on the literature and clinical practice experience. Following two rounds of discussion, 16 final recommendations were made, with the recommendation strength divided into recommend, suggest and not recommend.-Utilization of chest images and bronchoscopy1. Chest CT, rather than chest X-ray, is recommended for possible CAPA or CAPM patients to provide diagnostic evidence and localization for bronchoscopy to obtain microbiological specimens. A diagnosis of CAPA could not be made on the basis of positive signs on chest CT alone. Chest contrast CT or pulmonary artery CT (CTPA) is recommended in patients with probable CAPM.2. In the case of possible CAPA or CAPM, it is recommended that bronchoscopy and BALF collection for microbiological examinations be pereformed as soon as possible.-The selection strategies of microbiological examinations3. Microscopic examination, culture, GM testing and PCR for aspergillus Spp. of BALF are recommended in patients with probable CAPA. Fungal staining and culture of BALF are suggested for possible CAPM. Selected appropriate specimens for molecular biological detection are suggested in critically ill patients and possible CAPM.-Diagnostic critieria4. The revised ECMM/ISHAM consensus statement is recommended as the diagnostic criteria for CAPA and the Delphi consensus statement is recommended as the diagnostic criteria for CAPM.-Appropriate time for antifungal therapy5. Prophylactic therapy of CAPA with amphotericin B or its liposomes is suggested for patients with severe COVID-19, especially those with risk factors for CAPA.6. It is recommended to start the empirical anti-Aspergillus therapy as soon as possible for possible CAPA, and obtain the microbiological evidence for aspergillosis at the same time.7. Prophylactic therapy for CAPM is not recommended for severe COVID-19 patients.8. Early initiation of empirical therapy for possible CAPM is recommended, and microbiological evidence should be obtained at the same time.-Clinical applications for antifungal agents9.Voriconazole or isavuconazole are recommended as initial treatment for CAPA. Amphotericin B liposomes are suggested as the initial treatment for CAPM. Isavuconazole or posaconazole may be an option in patients with renal insufficiency or amphotericin B liposome intolerance/unavailability.10. In CAPA patients with tracheobronchitis, antifungal drug inhalation is recommended in addition to systemic antifungal medication.11. Combination therapy is not recommended as initial therapy for CAPA, but may be used as a salvage therapy strategy. Triazole or amphotericin B in combination with caspofungin or micafungin is recommended; whereas amphotericin B in combination with triazole is not recommended. For CAPM patients with extensive lesions, rapid progression or poor general condition, a combination of amphotericin B liposome with isavuconazole or posaconazole is suggested.-Response assessment and treatment duration12. It is recommended that treatment response be assessed comprehensively according to the clinical symptoms/signs, imaging and microbiological examination of patients. CAPA can be evaluated in combination with the dynamic change in serum GM.13. The recommended treatment duration of CAPA is at least 6-12 weeks. A total course of at least 3-6 months is suggested for CAPM, and the sequential treatment should be considered according to the response to 4-6 weeks of intravenous therapy.-How to adjust the anti-inflammatory therapy14. In patients with severe COVID-19 combined with possible or probable filamentous fungal infection, it is suggested that of anti-inflammatory therapy be stopped or reduced appropriately, taking into account of the severity of the infection and inflammation of the disease course. The combination of baritinib and/or tozzizumab based on glucocorticoids is not suggested in these patients.-How to treat the underlying diseases15. In patients with diabetes, strict glycaemic control is suggested. In patients with long-term use of glucocorticoids and/or immunosuppressants, it is suggested to reduce the intensity of immunosuppression. Granulocyte colony-stimulating factor is suggested to use to improve the circulating granulocyte levels in patients with granulocyte deficiency due to various causes.-When an operation should be considered16. In patients with CAPA, surgery is not recommended unless large blood vessels, pericardium, or chest wall are involved, or the patient has recurrent or massive hemoptysis. For CAPM patients, early surgical removal of lesions after diagnosis is recommended. Surgery is a high-risk procedure in patients with severe COVID-19, and a multidisciplinary team discuss is suggested.

Hydroalcoholic extract: A potential source from Azadirachta indica a Juss flower for DNA and BSA binding, scavenging, fungicidal, and larvicidal activities

AbstractThe present study focuses on the evaluation of the DNA and BSA binding capability, anti‐fungal, metal chelation, and larvicidal activity of extracts of Azadirachta indica A Juss flower. The different extracts of A. indica was prepared by soxhlet extraction using Petroleum Ether (PE), acetone (AC), and hydroalcohol (HA) as solvents. DNA and BSA binding ability of HA extract was calculated through the UV–Visible titrimetric method. The phytochemicals present in the extract show good binding ability towards DNA and BSA. H2O2 and NO scavenging activity were evaluated and HA extracts shows good antioxidant activity with IC50 values of 62 and 48 μg/mg, respectively. The metal chelating property was calculated, where HA shows good chelating ability with the IC50 value of 32.23 μg/mg. The antifungal activity of these extracts was evaluated against different fungal stains; among them, hydroalcoholic extract shows a higher zone of inhibition in all the fungal stains than Petroleum Ether and acetone extracts and acetone extracts (i.e., 50 μg/μL MIC). The potential extract (i.e., HA extract) was used to estimate the larvicidal activity of parasites. The % of mortality was evaluated through a dose‐dependent assay and it is confirmed that when concentration increases, the mortality rate increases. The correlation between metal chelating activity and total flavonoid and phenolic content was calculated and it was found that there is a good correlation (R2 – .743 and .902, respectively). These results proved that HA extract possesses biologically important phytochemicals that contribute to pharmaceutical and biological activities and could be a natural remedy for various real‐time applications.

A Case Report of Oral Ulcer in Rheumatoid Arthritis: A Diagnostic and Management Dilemma

Chronic oral ulcer in patients with rheumatoid arthritis is a diagnostic challenge in primary health care. The possible causes include the disease itself, side effect of treatment, malignancy and infections. This is a case of a 63-year-old woman with underlying rheumatoid arthritis presented with chronic solitary oral ulcer. She was on oral methotrexate (MTX) and high dose folic acid for five years with good compliance. Tissue biopsy showed marked inflammatory cells infiltration and nuclear hyperchromatism with no evidence of malignancy nor infection. The ulcer was resolved after completing antifungal treatment for six weeks following failure of improvement after empirical antibiotic, elimination of possible trauma and discontinuation of MTX. The fungal staining was negative. There are possibilities of false negative results, thus broadened diagnosis with repeated and further investigations are recommended in cases who did not respond to conventional treatment.