Rhino-orbital Mucormycosis in a COVID-19 Patient: The First Case in Malaysia

Mucormycosis is an aggressive and potentially fatal fungal infection caused by fungi of the order Mucorales. There has been an increase in the number of cases of rhino-orbital mucormycosis in people with COVID-19, particularly in India. Rhino-orbital-cerebral mucormycosis is the most common manifestation of mucormycosis associated with COVID-19. We report the first case of rhino-orbital mucormycosis in a diabetic patient with SARS-CoV-2 infection in Malaysia. The diagnosis of mucormycosis was confirmed by histopathological examination, but the fungal culture and PCR results were negative. He was treated with antifungal therapy and had extensive debridement. Treatment of mucormycosis requires a multidisciplinary approach that includes addressing underlying risk factors, effective antifungal therapy, and surgical debridement.

Chronic Laryngotracheal Granulomatous Disease Secondary to Sporothrix schenckii in an Immunocompromised Patient

Objective: To describe a rare presentation of laryngotracheal granulomatous disease secondary to sporotrichosis. Methods: The authors report a case of laryngeal sporotrichosis in an immunocompromised patient, with accompanying endoscopic images and pathology. Results/case: A 72-year-old immunocompromised female with a history of rose-handling presented with a year of hoarseness and breathy voice. Flexible nasolaryngoscopy showed diffuse nodularity; biopsy of the lesions demonstrated granulomatous inflammatory changes, and fungal culture grew Sporothrix schenkii. Long-term itraconazole treatment was initiated, with improvement in dysphonia and few residual granulomas on follow-up examination. Conclusion: When evaluating granulomatous disease of the airway, a broad differential including infectious or inflammatory etiologies should be considered, especially in immunocompromised patients. Adequate tissue samples should be collected to facilitate special staining. The current recommendations for laryngeal sporotrichosis include treatment with a prolonged course of itraconazole.

Clinical features of invasive fungal disease in children with no underlying disease

There is limited research into Invasive fungal disease (IFD) in children with no underlying disease. We undertook a retrospective study of children with IFD who did not suffer from another underlying disease, from June 2010 to March 2018 in Changsha, China. Nine children were identified. Eosinophil counts were elevated in six cases. The level of procalcitonin (PCT) was elevated in six cases. Fungal culture was positive in all patients, including eight cases of Cryptococcus neoformans and one case of Candida parapsilosis. 8.33 days following antifungal treatment, the body temperature of the eight patients affected by cryptococcal disease had returned to normal. Our study indicates that the primary pathogen in IFD was Cryptococcus neoformans in children who had no other underlying disease. Eosinophils can be considered to be indicators of cryptococcal infection. IFD in children with no other underlying disease has a satisfactory prognosis.

Molecular characterization and vegetative growth of pathogenic seed-borne fungus, Curvularia lunata of tomato and its in vitro control measures

Present studies were conducted to isolate and identify the seed-borne pathogenic fungus from the selected tomato variety through morphological and molecular techniques based on the sequencing of internal transcribed spacer (ITS) region of 18S rDNA. According to the colony and conidial features, the fungus was identified as Curvularia sp. The obtained ITS sequencing showed above 99% similarity with Curvularia lunata in the NCBI database. The sequence of the fungus was deposited in NCBI GenBank under the accession number: ITS, MH382879.1. Besides, the phylogenetic tree further confirmed the taxonomic position of the studied fungus. Growth characteristics of the fungus on nine different fungal culture media were evaluated, in which Honey peptone agar, Carrot agar, Potato sucrose agar, and Kauffman’s agar were found the most suitable. The maximum vegetative growth of the fungus was recorded at 30°C temperature and pH conditions. The bio-control potential of five different antagonists against the studied fungus was assessed, in which Trichoderma harzianum showed the better performance to restrict mycelial growth. Three ethanolic plant extracts were also evaluated, in which Lowsonia inermis L. exhibited above 60% mycelial growth inhibition of the fungus. Among three tested fungicides, Tilt 250 EC was found as an excellent fungicide to inhibit mycelial growth of C. lunata under in vitro conditions. Int. J. Agril. Res. Innov. Tech. 11(2): 124-132, Dec 2021

Sinonasal mycosis following transfrontal craniotomy in three dogs

CASE DESCRIPTION Three dogs were presented for investigation of chronic nasal discharge and epistaxis 141, 250, and 357 days after undergoing transfrontal craniotomy to treat an intracranial meningioma (2 dogs) or a meningoencephalocele (1 dog). CLINICAL FINDINGS CT findings were consistent with destructive rhinitis and frontal sinusitis in all 3 dogs, with results of histologic examination and fungal culture of samples obtained during frontal sinusotomy confirming mycotic infection. Frontal sinusotomy revealed fungal plaques covering a combination of bone and residual surgical tissue adhesive at the site of the previous craniotomy in all 3 dogs. Aspergillus spp were identified in all 3 dogs, and Chrysosporium sp was also identified in 1 dog. TREATMENT AND OUTCOME Surgical curettage was followed by antifungal treatment (topical clotrimazole in 2 dogs and oral itraconazole for 3 months in 1 dog). Nasal discharge improved in the short-term but recurred in all dogs 99, 118, and 110 days after frontal sinusotomy. One dog received no further treatment, 1 dog received an additional 8.5 months of oral itraconazole treatment, and 1 dog underwent 2 additional surgical debridement procedures. At last follow-up, 2 dogs were alive 311 and 481 days after frontal sinusotomy; the third dog was euthanized because of status epilepticus 223 days after frontal sinusotomy. CLINICAL RELEVANCE Sinonasal mycosis should be considered as a potential complication in dogs developing persistent mucopurulent nasal discharge, intermittent epistaxis, and intermittent sneezing following transfrontal craniotomy. The pathophysiology may be multifactorial, and potential risk factors, including use of surgical tissue adhesive in the frontal sinus, require further investigation.

Refractory feline sporotrichosis: a comparative analysis on the clinical, histopathological, and cytopathological aspects

ABSTRACT: Sporotrichosis is a chronic fungal infection caused by Sporothrix species. The occurrence of cases that are resistant to long-term treatment, especially in the nasal planum of cats, emphasizes the importance of studying its pathogenesis. The purpose of this study was to analyze and compare the inflammatory process of cutaneous lesions of feline refractory sporotrichosis to clinical aspects through cytopathological and histopathological examination. Moreover, the study included 13 cats with cutaneous lesions that had been resistant to itraconazole treatment for more than a year. Cutaneous lesions samples were collected for cytopathological, histopathological, and fungal culture analyses. Tissue fragments were processed and stained with hematoxylin-eosin (HE) and Grocott methenamine silver (GMS). Further, two clinical presentations had the highest occurrence: the localized cutaneous form in animals with good general condition and stable disease (n=9, 69.2%) and the disseminated cutaneous form in cats with poor general condition (n=4, 30.8%). In cats with refractory sporotrichosis, the nasal planum (84.6%) was the most common location of lesions. In the cytopathological study, cats with fewer than two lesions and in good general condition (n=9, 69.2%) showed absence or mild yeast intensity (up to 5 yeasts per field), lower intensity of macrophages and neutrophils, and higher intensity of epithelioid cells, lymphocytes, plasma cells, and eosinophils. On the other hand, (n=4, 30.8%) of the cats with disseminated sporotrichosis and a poor general condition had a marked intensity of yeasts, which were mostly phagocytosed by an increased number of macrophages and neutrophils. Of those animals with good general condition, the majority (n=6, 66.7%) had higher eosinophil intensity. In histopathology, malformed suppurative granuloma was the predominant type (n=9, 69.2%) in feline sporotrichosis lesions, followed by well-formed granulomas (n=4, 30.8%). Malformed granulomas showed mild to moderate fungal intensity (55.6%) in animals with good general condition and localized lesions while marked fungal intensity (44.4%) in cats with the disseminated form of the disease and poor general condition. Well-formed granulomas (n=4, 30.7%) had mild to moderate intensity of fungal load, and 75% of the animals with this type of granuloma had only one lesion and were in good general condition. Long-term itraconazole treatment in these cats with refractory sporotrichosis can keep the infection under control and localized lesions stable; however, fungus reactivation can occur, resulting in an exuberant and inefficient immune response.

First Report of Anthracnose Caused by Colletotrichum acutatum On Olive Fruits in Pakistan

Anthracnose symptoms on olive (Olea europaea) fruits cv. “Gamlick” were found in farmer orchards in Chakwal, Punjab, Pakistan (32° N and 72° E), with an average prevalence of 59%. Fruit symptoms start as thin, black, sunken lesions with a watery appearance that grow in diameter and coalesce into a large sunken soft zone. Lesions on mature fruit become noticeable in 5 to 6 days after infection, if temperatures are favorable (28°C). On the fruit lesion, orange conidial masses in dispersed or concentric circle arrangement can appear. Fragments (5 mm) were taken from the margins of fruit lesions and surface-sterilized with 70% ethanol (1 min) and 1% NaClO (2 min), cleaned with sterile purified water, blotted dry, and plated on potato dextrose agar (PDA) in Petri dishes. The petri plates were incubated at 27°C. A fungus was consistently isolated, and thirty-five isolates were characterized. Aerial mycelia from olive isolates Colonies were compact, initially white or cream white, then grey, and eventually dark grey, with conidium masses forming in the middle. Mycelium is branched, septate, and hyaline. Conidia are hyaline, aseptate, fusiform, or often cylindrical, with obtuse apices and tapering bases. Their mean size was 8.5µm in length and 3.0 µm in width. Based on morphological features, the fungus was tentatively identified as Colletotrichum acutatum (Agosteo G.E., 2010). The identification was confirmed by amplification and sequencing of a representative isolate's internal transcribed region (ITS), Beta- tubulin region (TUB2), Actin region (ACT), and Glyceraldehyde 3-phosphate dehydrogenase region (GAPDH) with the primers ITS1/ITS4 (Gardes & Bruns 1993), TUB4/TUB5 (Woudenberg et al. 2009), ACT1/ACT3 (Carbone & Kohn 1999) and GDF1/GDR1 (Guerber et al. 2003). BLAST analysis revealed 100% identity for ITS, GAPDH and ACT and 99% identity for TUB, between the sequences of the olive fruit isolate (GenBank Accessions MW647502, MZ436968, MZ714412 and MW810331, respectively) and sequences of C. acutatum reference isolates (GenBank Accessions GO613492, KF975660.1, MT274752.1 and MH547616 respectively). Phylogenetic analysis based on ITS, GAPDH and TUB regions of the olive fruit isolates and reference isolates of various Colletotrichum species using the MEGA X software program confirmed the isolate from olive was C. acutatum. The fungal isolate was deposited as a living culture in the Barani Agricultural Research Institute's fungal culture collection center (BACA.9381). Pathogenicity tests were conducted with this isolate by placing a 20 µl drop of a conidial suspension (3 × 107conidia ml−1) on five healthy olive cv. Gemlik fruits. As a control, five non-inoculated olive fruits were used. Fruits were placed at a temperature of 27°C with artificial light and a photoperiod of 12 hours. Anthracnose symptoms developed only on inoculated fruits after seven days of inoculation. The fungus was re-isolated from symptomatic fruits, and its identity was confirmed through morphological characteristics, thus verifying Koch's postulates. To the best of our knowledge, this is the first report of C. acutatum infecting olive fruits in Chakwal region of Pakistan.

Investigating Factors of False-Positive Results of Aspergillus Galactomannan Assay: A Case–Control Study in Intensive Care Units

Background: Studies on false-positive galactomannan (GM) enzyme immunoassay (EIA) results and treatment for critically ill patients are scarce.Objectives: The study aimed to determine the false-positive rate of GM-EIA and to probe the risk factors of false positivity among patients in the intensive care units (ICUs).Methods: A case–control approach was conducted to review adult patients who had at least one GM-EIA result and were admitted to the ICU. Those who had no fungal culture were excluded. The clinical characteristics and critical care between patients with false-positive and true-negative GM index (GMI) were compared.Results: Of 206 patients enrolled and with GM-EIA results, 20 (9.7%) were considered to have false-positive antigenemia, including 9 in bronchoalveolar lavages (BAL) and 11 in serum. A total of 148 (71.8%) were true-negatives. After paired grouping of 1:4, factors researched in the previous studies showed no significant difference. However, compared with the true-negatives, patients with positive GM test results but were incompatible with the diagnosis of invasive aspergillosis were more prone to the risk of false positivity due to the use of colistin inhalation. It seemed to be the only factor that significantly increased the risk of false positivity after multivariate analysis (adjusted odds ratio, 35.68; 95% CI, 3.77–337.51, p = 0.002).Conclusions: Colistin inhalation treatment may contribute to false-positive GM-EIA results. The positive GMI among patients receiving colistin nebulization should be interpreted with caution.

Spectrum of dermatophytes in a tertiary care centre in Northern India

Dermatophytes is major public health challenge in many parts of the world, mainly in developing countries due to poor housing facilities, high population per capita, and poor sanitary conditions. Early diagnosis and identification is must for preventing and early treatment of dermatophytosis. Also, some studies suggest that in prepubescent children there is an inadequate amount of fungi inhibiting fatty acids synthesized predisposing them to dermatophytic infections. Reduction in the synthesis of these fungistatic triglycerides in sebum premenopausal women is also seen predisposing them to infection by dermatophytes. However, low socioeconomic status along with illiteracy and overpopulation has been a main predisposing factor to dermatophytic infections in developing parts of the world. The incidence also been increased due to the rise in the number of immunocompromised patients and considerable use of broad-spectrum antibioticsEarly finding of infection is must for prevention and early management of dermatophytosis. Dermatophytes enter keratinized tissue via keratinases, which produce a dermal inflammatory response causing burning, itching and rednessTo determine prevalence of species of dermatophytes. This Study was conducted on 334 samples i.e. skin scrapings, nail clippings, and hair for fungal culture in the Mycology laboratory over a period of one year extending from December 2019 to December 2020. Specimens were cultured on modified Sabouraud's dextrose agar media containing antibiotics and incubated at 25°C and 37°C for a period of 4 weeks. Isolation and identification of various species of dermatophytes were done. A cross-sectional study was conducted on patients who came to our hospital in the department of dermatology or were referred to the department of microbiology over a period of one year extending from November 2019 to November 2020. Microbiological tests of suspected patients included potassium hydroxide (KOH) mount and fungal culture examination. Cases with culture-positive results were correlated with clinical diagnosis. In the study total of 334 samples (skin scrapings, nail clippings, and hair) were received for fungal culture in the microbiology laboratory during the study period. Samples obtained were cultured on modified Sabouraud's dextrose agar (SDA) media containing antibiotics and incubated at 25°C and 37°C for a period of 4 weeks. Species identification was performed based on colony's morphology, finding of the teased mount by using lactophenol cotton blue stain (LCB) and slide culture, and also with urea hydrolysis test as seen.The Study was performed on 334 samples received from the department of dermatology for fungal culture. Fungal elements were seen in 31% of cases and were isolated in 30% of cases. In these culture-positive cases, dermatophytes were reported in 90% cases, Candida species in 4%, and another fungus was reported in 6% cases. Trichophyton species is most commonly isolated (27.6%). Microsporum and Epidermophyton species were isolated in 5.1% cases. T. mentagrophytes was the most common fungal isolate among all the culture-positive cases.

The clinical type and etiological agents of superficial dermatophytosis: A cross sectional study

Dermatophytosis are fungal infections caused by three genera of fungi that have the unique ability to invade and multiply within keratinized tissue (hair, skin, and nails). Although dermatomycoses are globally distributed, the endemic and most prevalent species of dermatophytosis differ strikingly from one geographic locality to another. Changing trend has been noticed in last few years with dermatophytic infections presenting as chronic, treatment unresponsive and recurrent. Also various microscopic and fungal culture studies have shown shift in identification of causative fungal species in recent years. Numerous studies have been done on the occurrence of dermatophytes in various parts of our country illustrating the range and changing pattern of fungal infection as well as causative fungal species. Total number of 150 patients attending outpatient department of our hospital who were clinically diagnosed as having superficial dermatophytosis were enrolled into the study. Patients were carefully screened as per inclusion and exclusion criteria and then enrolled in the study. Samples were taken from all the patients and examined for KOH direct microscopy and sent for fungal culture on Sabouraud’s Dextrose Agar as well as on Dermtophyte Test Medium. Results were then analyzed using standard statistical methods. Out of total 150 patients, 101 were males and 49 were females. Most common age group was 21-30 years (37.3%). 58 patients (38.7%) showed positivity to KOH microscopy as well as fungal culture. Additionally 25 more samples demonstrated positivity to KOH microscopy (total 83 patients) but negativity to culture, while 9 patient samples were positive to culture but negative to direct microscopy. Predominant fungal species isolated on culture was Trichophyton mentagrophytes (50.7%) while next common species isolated was T. tonsurans (29.9%). No significant association was found between dermatophyte isolate on culture and clinical type. Trichophyton mentagrophytes and Trichophyton tonsurans were the most common species isolated among subjects with Tinea faciei, Tinea cruris and Tinea corporis. The study showed a male preponderance and T. corporis was the commonest clinical type found. Majority of patients were in the 3rd decade and came within a duration of 1 month to 6 months of getting an infection. In patients diagnosed with tinea corporis, tinea cruris, tinea pedis and tinea manuum, T. mentagrophytes was the most predominant species isolated.