Czapek-Dox Agar Research Articles

Evaluation of hydrocarbon degrading potentials of cellulolytic edaphic borne fungi

This study assessed the hydrocarbon degradative potentials of cellulolytic fungi cultured from top soil samples within Benin City, Edo State. Soil samples were collected from 3 sites; a farmland, a fallow plant covered land, and municipal dumpsite. Physicochemical parameters of the soil samples were based on standard methods. Standard cultural and molecular techniques were used to isolate and identify fungi present in samples. Cellulolytic activity of fungal isolates was determined with Czapek-dox agar supplemented with 0.1% sodium carboxymethyl cellulose (CMC). Cellulolytic fungi were subsequently screened for the ability to utilize blended petroleum as sole carbon source. Further, two isolates having the highest optical density were used for growth profile study as axenic culture and in consortium. The growth profile study assessed pH, optical density, total petroleum hydrocarbon content and fungal dry mass. Results showed that the organic carbon value of the top soils ranged from 4.14 g/kg ± 0.55 to 8.56 g/kg ± 0.05. All collected soil samples were sandy, while total hydrocarbon content ranged from 0.38 g/kg ± 0.04 to 1.13 g/kg ± 0.01. The mean fungal counts were highest in dumpsite soils and ranged from 4.0 × 103 cfu/g ± 1414 to 7.0 × 103 cfu/g ± 1003. Eight fungal isolates with cellulolytic properties were identified from the soil samples. Trichosporon asahii and Curvularia bothriochloae had highest (p < 0.05) OD when grown on blended petroleum and were used for growth profile studies. OD was highest (p < 0.05) in C. bothriochloae (0.372 ± 0.0005) relative to T. asahii and the consortium at 15 d. THC reduced significantly (p < 0.05) in both axenic cultures and the consortium after 15 days, while fungal biomass was highest (p < 0.05) in consortium relative to axenic cultures after 15 d. This study showed hydrocarbon degrading capabilities of cellulolytic fungi from different top soils.

Production of Citric Acid from Different Aspergillus species Obtained from Soil

The genus Aspergillus is considered highly important in the production of various types of enzymes and organic acids. Aspergillus species produce organic acids such as citric acid, itaconic acid, and malic acid, which are one of the most important alternate techniques for chemical processes. Citric acid is an essential primary agricultural product that is extensively used in the world. It acts as an intermediate in the tricarboxylic acid cycle (citric acid cycle) during the oxidation of carbohydrates to carbon dioxide. Citric acid has an important role in industries involving the production of food products, beverages, pharmaceutical and agricultural products, detergents, and cosmetics. The production of citric acid, a crucial agricultural commodity with widespread applications in various industries including food, beverages, pharmaceuticals, agriculture, detergents, and cosmetics, heavily relies on microbial fermentation. The great demand for citric acid due to its wide industrial applications and less toxicity. Citric acid can be produced using less expensive substrates that are renewable too. In this work, Aspergillus species was isolated from the soil source, collected in different localities of Chhatrapati Sambhajinagar. The total 90 different isolates are isolated among these the 20 different Aspergillus species are isolated and pure culture by using potato dextrose agar (PDA) Czapek dox agar medium and study the effect of different media on growth and sporulation of different Aspergillus species and the quantitative analysis of citric acid production was done by titration method with help of NaOH and phenolphthalein indicator. The main reason for the study increases is the large number of applications that can be found for citric acid, primarily in the food and pharmaceutical industries

Role of Copper Ions in Resistance of Modern Polymer Composite Materials to Fungal Damage

Resistance of polymer composite materials to biodamage is one of the pressing problems of our time. Incorporation of Cu2O (I) in the composition of a polymer composite based on the ED-20 epoxy resin increases its biocidal properties. Under conditions of mineral and organic contamination, the area of the samples affected by micromycetes was found to decrease with increasing concentration of dispersed particles in the composite. The affected area of the samples filled with the particles encapsulated in polylactide was 1.5 times smaller than that of the composites filled with non-encapsulated particles. Сopper oxide had a toxic effect on the Aspergillus niger strain dominant on the surface of the samples, causing a decrease in the average radial growth rate on the Czapek-Dox agar medium and in the biomass weight concentration during the growth of micromycetes in a liquid medium compared to the variant without Cu2O.

L-Asparaginase producing ability of Aspergillus species isolated from tapioca root soil and optimized ideal growth parameters for L-Asparaginase production

This research was designed to isolate the predominant L-asparaginase-producing fungus from rhizosphere soil of tapioca field and assess the suitable growth conditions required to produce maximum L-asparaginase activity. The Aspergillus tubingensis was identified as a predominant L-asparaginase producing fungal isolate from 15 isolates, and it was characterized by 18S rRNA sequencing. The L-asparaginase-producing activity was confirmed by pink color zone formation around the colonies in modified Czapek Dox agar plate supplemented with 1% L-Asparagine. The optimal growth conditions required for the L-asparaginase production by A. tubingensis were optimized as pH 6.0, temperature 30 °C, glucose as carbon source, 1.5% of L-Asparagine, ammonium sulphate as nitrogen source, rice husk as natural L-Asparagine enriched source, and 8 days of the incubation period. The L-Asparaginase activity from A. tubingensis was excellent under these optimal growth conditions. It significantly used rice husk as an alternative to synthetic L-Asparagine. As a result, this may be considered a sustainable method of converting organic waste into valuable raw material for microbial enzyme production.

Etiology and Antifungal Sensitivity Test in Otomycosis Caused by Candida Sp

Otomycosis is a common fungal infection of the external auditory meatus frequently diagnosed in otolaryngology outpatient clinics. Resistance to antifungals is currently a significant concern, with intrinsic and acquired resistance increasing among isolates that cause fungal infections. The purpose of this research was to identify Candida species causing otomycosis and determine the pattern of antifungal susceptibility among these Candida species. A prospective study was conducted in the Margono Soekarjo General Hospital and Department of Microbiology, Faculty of Medicine, Universitas Jenderal Soedirman Purwokerto, Indonesia, from April – September 2022. Forty-seven (47) clinical samples of otomycosis were collected from 41 patients and then isolated bedside on fungal culture media and was prepared on an object glass for direct microscopic examination of the specimens. Fungal identification was performed using 10% potassium hydroxide (KOH) to observe fungal elements. Samples were cultured on Saboraud dextrose agar (SDA) media with chloramphenicol and Czapek dox agar. The Germ Tube Test was used to identify Candida while yeast-specific identification and antifungal susceptibility assay using a rapid commercial kit was applied for specific identification of the fungus. Antifungal susceptibility patterns were obtained using the Integral System Yeast Plus (ISYP) media pack. Candida parapsilosis was the most prevalent Candida species discovered in this study, accounting for approximately 41.66%, which was followed by Candida tropicalis (25%) and Candida krusei (12.5%), whereas Candida albicans only accounted for 4.1% of the specimens. All Candida species were sensitive to flucitosine and ketoconazole, whereas the voriconazole sensitivity rate reached 96%. This study concludes that Candida parapsilosis is the most prevalent species of Candida in otomycosis, and that all Candida species are sensitive to Flucitosine, ketoconazole, and voriconazole.

Contamination of Armenian Red Pepper by Microfungi and Aflatoxin B1

Key words: paprika, hot red pepper, filamentous fungi, aflatoxin B1, strains, contamination The aim of this study was to investigate the contamination of ground red pepper with mycotoxigenic fungi. During 2019-2020 we have analyzed 45 samples taken from different regions of the republic. 13 species of filamentous fungi, belonging to five genera were isolated from ground pepper. Such spices as Aspergillus niger, A. flavus, A. fumigatus var. griseobrunes, A. ochraceus, Penicillium lanosum и Absidia corymbifera were most frequently isolated from red pepper samples on Czapek-Dox agar medium. A chromatographic analysis of 17 samples of ground red pepper was carried out for the presence of aflatoxin B1. As a result, Aflatoxin B1 had been detected in 9 samples. Aflatoxin concentrations in paprika and hot red pepper ranged from 10 to 50 μg/kg. The aflatoxin content in red pepper samples did not depend on its pungency. Other mycotoxins have not been identified during the present study. These findings indicate the risk possibility of human exposure to mycotoxins through the consumption of dry rep pepper.

Identification of Previously Unrecorded Bacills, Serratia, and Mucor Strains Isolated from Yogurt

(1) Background: Foodborne illnesses are recognized as a significant threat to public health and the economy in both developed and developing nations. The safety of foods containing microorganisms has consequently become a major worldwide concern. One of the most frequent causes of food deterioration in the world is microbial contamination. (2) Methods: Yogurt containers that were bought commercially in Riyadh, Saudi Arabia during their validity period contained four different species of bacteria and one type of fungus. Using nutritional agar and Czapek-Dox agar medium, the bacteria and fungi were isolated. The isolates of the fungi and bacteria were identified using a scanning microscope. The isolates were further identified and classified for molecular evolutionary analyses using the 16S rRNA and ITS sequences from the bacteria and fungi, respectively, in conjunction with the universal primers 27F, 1492R, ITS1F, and ITS4R. (3) Results: A total of 131 separate strains were identified from 12 yogurt samples based on their phenotypic characteristics. In total, 79 isolates (60.3%) consisted of Serratia marcescens, Bacillus subtilis, and Mucor circinelloides, with 52 isolates (39.7%) being Bacillus cereus. While the cells of Bacillus and Serratia are shaped like rods, the sporangia of Mucor are large, round, and black. Each strain was identified by its accession number, which were as follows: MK590996.1: B. cereus; MK591144.1: B. subtilis; MK591002.1 and MK591014.1: S. marcescens; and MK559692.1: M. circinelloides. The maximum identification was found to be between 98.64 and 100% when BLAST was used to compare the sequences to the NCBI GeneBank database. (4) Conclusions: Genus and species identification was performed using the similarity score values. Yogurt products containing high concentrations (39.7%) of Bacillus cereus isolates carry a significant risk of health hazards due to the potential for spreading pathogenic bacteria to humans.

Cultural Characteristics of Rhizoctonia solani Causing Sheath Blight of Rice

Sheath blight of rice caused by Rhizoctonia solani Kuhn is emerging as a very destructive disease causing heavy yield losses. The present study was conducted at Department of Plant pathology, JNKVV, College of Agriculture, Rewa, (M.P.) with eight culture media viz., Potato Dextrose Agar (PDA), Czapek’s Dox Agar (CDA),Richards medium (RMA), Corn Meal Agar (CMA), Oat meal agar (OMA), Rice leaf extract agar (30%),Rice seed extract agar (10%) and Rice seed extract (10%)+ (2%) sucrose were evaluated against R. solani. It can be concluded that Potato dextrose agar (88.0 mm) and Czapek’s dox agar (87.7 mm) medium supported maximum mycelial growth of R. solani isolated from rice. Whereas, least mycelial growth (75.8 mm) was observed in Rice seed extract agar and Richards agar medium. Mycelial growth was excellent fast, abundant and off white on Potato dextrose agar. Whereas it was fast, abundant and on Czapek’s dox agar. Sclerotia of the fungus initiated within 3 - 4 days in different culture media. Maximum number of sclerotia was produced in Oat meal agar (54.0) closely followed by Potato dextrose agar (51.2) and Czapek’s dox agar (44.6). Maximum weight of 10 sclerotia was recorded in potato dextrose agar (1030.6 mg) followed by Czapek’s dox agar (920.3 mg) and Oat meal agar (884.7 mg). Maximum size of sclerotia were formed in Potato dextrose agar followed by Rice seed extract agar and Corn meal agar.

Screening of Aflatoxin Production by Aspergillus flavus Isolates from Petroleum-contaminated Soil

Fungi are eukaryotic, heterogeneous, unicellular to filamentous, spore-bearing, and chemoorganotrophic organisms which lack chlorophyll. This present study was carried out to isolate and identify fungi from petroleum-contaminated soil. Several fungal genera which included Rhizopus spp., Mucor spp., Penicillium spp., Rhizoctonia spp., Aspergillus spp., Alternaria spp., and Cladosporiumspp. were isolated using potatoes dextrose agar, Czapek-Dox Agar, and Aspergillus flavus Differentiation Agar culture media that comparable with co-amoxiclav (1g) and chloramphenicol to prevent the growth of any bacteria. The direct plate and serial dilution agar plate methods were used for the isolation of fungi. Based on results, Aspergillusand Mucor spp. were the most predominant genera and had the highest number of colonies in the soil samples. In this investigation, seven out of 27 soil samples were morphologically (macroscopically and microscopically) identified, such as A. flavus. Aflatoxigenicity of A. flavus was detected using characteristics in Aspergillus differentiation agar and colony fluorescence on exposure to ultraviolet light. Moreover, molecular approaches were used for the detection of aflatoxigenic of the A. flavus isolates. Three structural (aflD, aflO, and aflP) and one regulatory (aflR) gene of the aflatoxin gene cluster of A. flavus were targeted for amplification by the polymerase chain reaction method. The aflatoxigenic of all six A. flavus isolates was detected molecularly which contained two structural (aflD, aflP)genes out of three structural genes, while there was no specific amplification of the aflO gene in the fourth, fifth, and sixth A. flavus which issimilar to the aflR gene in the first and second A. flavus.

Growth and morphological characteristics of some pyrophilous discomycetes in culture

Three pyrophilous discomycetes species (Anthracobia maurilabra (Cooke) Boud., Pyronema domesticum (Sowerby) Sacc. and Tricharina praecox (P. Karst.) Dennis) have been cultivated and studied in vitro. Cultures were obtained from fresh apothecia сollected in the Holosiivskyi National Nature Park (Kyiv, Ukraine). The culture growth and morphological characteristics of the studied fungi on different media (beer wort agar, Czapek Dox agar, potato-dextrose agar) were analyzed. All investigated species can quite easily grow under laboratory conditions on different tested nutrient media. Potato-dextrose agar was the most suitable medium for the enhancement of radial growth and the best expresses all the phenotypes of the colony of studied fungi. Macro- and micromorphological descriptions of all fungal colonies and illustrations are provided. The micromorphological analysis showed that common to the mycelium of all studied species of fungi was the presence of numerous drops of oil in the hyphae, anastomoses like T. praecox, а net- like structure of A. maurilabra similar to nematode capture hook. Beside this, A. maurilabra and P. domesticum formed the sexual stage under experimental conditions. Forming fruiting bodies of A. maurilabra in culture has not been reported before. Moreover, P. domesticum was found to form abundant dark brown sclerotia on potato-dextrose agar and Czapek Dox agar. Possible pigment composition in the P. domesticum sclerotia is discussed based on the Raman spectroscopy study, performed on this genus for the first time. The established cultural characteristics can be useful for taxonomic identification of fungal species and for pure quality control of mycelial cultures during their introduction, preservation and future potential applications in biotechnological areas.

PREVALENCE OF (Aspergillus niger) IN OTOMYCOSIS AT PROF. DR. MARGONO SOEKARJO HOSPITAL

Otomycosis is a chronic fungal infection with high prevalence. Environmental factors dan predisposing factors can influence a patient’s vulnerability to otomycosis. According to studies conducted in several country, Aspergillus niger is the common fungus that causes otomycosis. This study aims to determine the prevalence of Aspergillus niger in patients at Prof. dr. Margono Soekarjo Hospital, who was clinically diagnosed with otomycosis. The study was done at the ENT Poly of Prof. dr. Margono Soekarjo Hospital and Microbiology Laboratory of Medicine Faculty, Jenderal Soedirman University between August 2022 to Januari 2023 as a descriptive observational study. According to the inclusion criteria, there were 42 subjects and 46 samples that ENT specialists carried out with sterile flocked swabs, the isolated the samples bedside on SDA Chloramphenicol, Czapek Dox agar, and object glass. Samples were observed macroscopically and microscopically with 10% KOH and LPCB staining. Patients with Aspergillus niger infection in otomycosis at Prof. dr. Margono Soekarjo Hospital , are represented by nine (19,56%) out of the 46 samples. The gender with the highest sex precentage is female (55,56%), folllowed by the age range of 26 – 35 years (44,45%). The prevalence of Aspergillus niger as a cause of otomycosis in Prof. dr. Margono Soekarjo Hospital is 19,56%, the most in females and the age group 26 – 35 years.

The Mechanism of Ochratoxin Contamination of Artificially Inoculated Licorice Roots.

Ochratoxin (OT) contamination of medicinal herbs is a serious threat to human health. This study was performed to investigate the mechanism of OT contamination of licorice (Glycyrrhiza sp.) root. Licorice root samples were cut into eight parts, which were placed separately on sucrose-free Czapek Dox agar medium, inoculated with the spores of ochratoxigenic Aspergillus westerdijkiae. After incubation for 10 and 20 days, the OT contents of the samples were determined by high-performance liquid chromatography, and microtome sections prepared from the samples were analyzed by desorption electrospray ionization tandem mass spectrometry, to visualize OT localization. The same sections were further examined by light microscopy and scanning electron microscopy, to investigate the path of fungal mycelial penetration of the inner roots. OT concentrations tended to increase from the upper- to the middle-root parts. OTs were located in cut areas and areas of cork layer damage; they were not present in the undamaged cork layer, indicating that the structure of this layer prevents OT contamination of the licorice root.

Culture Media Options for Growth and Morphological Characterisation of Cercospora coffeicola Affecting Coffee in Zimbabwe

Cercospora leaf spot is fast turning into a critically important diseasein Zimbabwe. The disease is caused by Cercospora coffeicola whichsignificantly reduces productivity and quality of coffee. Disturbingly,optimum sporulation of Cercospora coffeicola in culture remains a limitingfactor for microbial analysis and quantitative studies of Cercosporaleaf spot. Faced with this challenge, an in-vitro study was conducted atCoffee Research Institute, Manicaland, Zimbabwe to examine growth ofCercospora coffeicola in different nutrient media and to determine the bestmedia for Cercospora coffeicola analysis. Six nutrient media were assessed(corn meal agar, oat meal agar, Czapek Dox agar, malt extract agar, yeastextract agar and potato dextrose agar) for the growth of Cercosporacoffeicola. The laboratory-based experiment was duplicated, laid out ina Completely Randomized Design, replicated three times and based onCercospora coffeicola nutrient inoculation. Data were collected on radialgrowth, colour and texture of mycelium at 3 and 6 days after inoculation.There were significant differences (p < 0.05) in the growth of Cercosporacoffeicola in media after 3 and 6 days. Malt extract agar had the greatestradial growth (34 mm and 32 mm) of Cercospora coffeicola for trials 1and 2 respectively, whilst the least growth was in the oat meal agar (14.2mm and 15.7 mm) for trials 1 and 2 respectively. There were variations incolour and texture of mycelium with malt extract agar, potato dextrose agarand oat meal agar associated with darker colours and rough texture whilesmooth white mycelia were found in corn meal agar. After considering allnutrient media, malt extract agar was found to be the best media for thegrowth of Cercospora coffeicola in-vitro. On the basis of our findings, theauthors recommend the use of malt extract agar as the primary media foridentification and characterisation of Cercospora coffeicola.

Solation, Screening and Optimization for Laccase production by Fusarium spp. under submerged fermentation

Laccases are oxidases with broad substrate specificity and ability to oxidize various phenolic and nonphenolic compounds. This study was carried out to isolate and characterizes laccase producing fungi from the environmental samples. Soil samples were collected from different locations. Suspensions of the samples (1g in 10 ml sterile physiological saline) were serially diluted, inoculated onto Czapek Dox Agar (CZA) containing antibiotic chloramphenicol and incubated for 7 days at 30 o C. The fungal isolates were characterized macroscopically and microscopically. Screening was done using Czapek-Dox agar was supplemented with 0.1% tannic acid, 0.5% Bromophenol blue, 0.05 ml in 25 ml of Methanol Guaiacol the pure culture of the single colonies from the spread plate method were inoculated on the media and incubated at room temperature for 5-6 days. After 5-6 days the plates were checked for clear transparent zone.

Biological control of cabbage head rot (Sclerotinia sclerotiorum) by Coniothyrium minitans

BackgroundSclerotinia sclerotiorum is the causal agent of white mould, stem and head rot diseases on a wide variety, including cabbage in the field and storage. Control of this pathogen by using commercial disease management methods is extremely difficult. Therefore, this study was performed to identify the potential biological control agent.ResultsThe antagonist fungal isolates, a mycoparasite of S. sclerotiorum, were isolated and described as KET Cm1, PAL Cm 2 and TDK Cm 3. They were subjected to morphological and microscopic observation. The growth of antagonist fungal isolates on different media revealed that PDA and half-strength Czapek dox agar medium showed the maximum mycelial growth (28.67 and 28.33 mm) and pycnidial production in the PDA medium. The studies on pH and temperature found that growth and sporulation of the antagonist fungal isolates were optimum at pH 5.5 (34 mm) and 20 °C. The molecular confirmation of antagonist fungal isolates was carried out using ITS 1 and ITS 4 primers, which target 18S-28S rRNA gene fragment. The isolates were partially sequenced and showed that more than 99% identity with Coniothyrium minitans already exist in the NCBI database. The accession numbers (MW093134, MW111282 and MW136938) were obtained for all the three isolates and also confirmed using specific primers Cm sp 1F and Cm sp 1R. Among the tested isolates, the antagonistic activity of C. minitans TDK Cm 3 showed the maximum mycelial growth inhibition of 48.8% S. sclerotiorum and was used for further studies. Penetration and formation of appressoria on the hyphae of S. sclerotiorum by C. minitans isolate TDK Cm 3 were observed through the scanning electron microscope. The ethyl acetate fractions of TDK CM 3 isolate were evaluated against S. sclerotiorum by agar well diffusion assay, paper disc assay and poison food technique in vitro. Through GC–MS analysis, the effective antifungal compounds were identified.ConclusionThe present study focused on the identification and characterization of effective Coniothyrium isolates as eco-friendly, integrated disease management strategies against cabbage head rot disease.

P205 Uncommon Aspergillus species encountered in human infections

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objective Aspergillus species are ubiquitously found in environment and have propensity to infect both immunocompromised as well as immunocompetent individuals. Though the most common species implicated in human illness include Aspergillus fumigatus and A. flavus, many uncommon Aspergillus species are increasingly being reported. Moreover, these Aspergillus species have the potential to afflict varied organ systems and can thereby, be implicated in a wide array of infections. Here, we describe three patients infected by rare Aspergillus species viz. A. sclerotium, A. cristatus, and A. japonicum causing brain abscess, corneal ulcer and chronic dacrocystitis.MethodsUncommon Aspergillus species were isolated from clinical samples (nasal scraping, corneal scraping, and brain biopsy). Samples were inoculated on Sabouraud dextrose agar (incubated at 37°C and 25°C) and brain heart infusion agar (incubated at 25°C), followed by slide culture and lactophenol cotton blue (LCB) mount. The isolates were further subjected to molecular identification using polymerase chain reaction (PCR) targeting ITS region, followed by sequencing. Demographic details, clinical characteristics, risk factor, and management profile associated with these Aspergillus species were evaluated. We conducted systematic review using the search terms ‘A. sclerotium’ AND ‘humans’, ‘A. cristatus’ AND ‘humans’ and ‘A. japonicum’ AND ‘humans’.ResultsWe isolated three newer Aspergillus species from nasal scraping, corneal ulcer and brain biopsy. Direct microscopy using potassium hydroxide-calcofluor mount from all these clinical samples showed hyaline septate hyphae in background of pus cells. After 3-5 days of incubation, yellow-green mycelia were observed on the obverse in all the isolates. However, their microscopic features did not relate with the typical lactophenol cotton blue (LCB) picture of A. flavus, owing to which these were sub-cultured on Czapek Dox agar. After 4-5 days of incubation, LCB of these isolated again failed to divulge any specific picture and these isolates were subjected to molecular identification.Nasal scraping from 31-year-old immunocompetent male who presented with post-traumatic chronic dacrocystitis and recurrent orbital cellulitis yielded A. japonicum. He responded well with empirical antibiotics and oral voriconazole.Corneal scrapings from 56-year-old immunocompetent male who presented with post-traumatic blurring of vision yielded A. cristatus. He was treated with nystatin and voriconazole with good response.Brain biopsy from 45-year-old male with hypertension and subarachnoid hemorrhage, who presented with fever, headache, vomiting and periorbital swelling yielded A. sclerotium. He was treated with amphotericin B and had favorable outcome.The systematic literature review didn't yield any results of infection caused by these species in humans.ConclusionThis is the first report of isolation of A. sclerotium, A. japonicum, and A. cristatus from human tissue, though these fungi have been isolated from ambient air and environment. The present report highlights the emergence of uncommon Aspergillus species causing invasive infections even in immunocompetent patients and the requisite of molecular modalities to aid in identification of these rare, emerging species.

First report on the metabolic characterization of Sterigmatocystin production by select Aspergillus species from the Nidulantes section in Foeniculum vulgare.

Spices are typically grown in climates that support the growth of toxigenic fungi and the production of mycotoxins. The Aspergilli described in this study, as well as the sterigmatocystin (STC) detected, are causes for concern due to their potential to induce food poisoning. One of the most well-known producers of the carcinogenic STC is Aspergillus nidulans. This research explores the occurrence of STC-producing fungi in Foeniculum vulgare, a spice that is marketed in India and other parts of the world. This innovative study details the mycotoxigenic potential of five Aspergilli belonging to Section Nidulantes, namely Aspergillus latus (02 isolates), Emericella quadrilineata (02 isolates), and Aspergillus nidulans (01 isolate), with respect to STC contamination. These five isolates of Aspergilli were screened to produce STC on yeast extract sucrose (YES) medium in a controlled environment with regard to light, temperature, pH, and humidity, among other variables. The expression patterns of regulatory genes, namely, aflR, laeA, pacC, fluG, flbA, pksA, and mtfA were studied on the Czapek–Dox agar (CDA) medium. STC biosynthesis by the test isolates was done in potato dextrose broth (PDB) under optimum conditions, followed by the extraction and purification of the broth using ethyl acetate. High-performance liquid chromatography (HPLC) with an ultraviolet (UV) detector was utilized to detect compounds in eluted samples. F. vulgare contains Aspergilli that have been shown to have mycotoxigenic potential, which can accumulate in the spice during its active growth and thereby cause the elaboration of mycotoxins.

PRODUCTION OF FUNGAL CELLULASE ENZYMES AND THEIR APPLICATIONS

Cellulose is the most common organic polymer. The enzyme cellulase may break it into smaller sugar components, such as glucose subunits, which can be utilized to make ethanol, organic acids, and other compounds. Cellulases are a set of enzymes that have applications in various industries, including food, textiles, pharmaceutical, detergent, biofuels, and paper, making them the third-largest industrial enzyme utilized worldwide with rising demand. Cellulolytic fungi were first screened by exposing them to Carboxyl Methyl Cellulose agar (CMC), a selective media for cellulolytic bacteria. Further testing can be done on Czapek-Dox agar medium supplemented with 1% CMC. The cellulase production was significantly improved by optimizing many parameters such as pH, temperature, and substrate concentration.

MOLD DIVERSITY OF WREATHED HORNBILL (Rhyticeros undulatus) NEST IN MOUNT UNGARAN

Wreathed Hornbill (Rhyticeros undulates) is known to build nests in three cavities where they managed to live and breed. This edifice is predicted to contain various molds needed to maintain micro-environmental steadiness. This study was aimed to identify molds diversity in the Wreathed Hornbill’s nest, using samples collected from empty structure with no bird activity. The samples were obtained from the Kalisidi and Nglimut observation stations on two occasions, i.e., in 2016 and 2017. Furthermore, the samples comprised cover soil, wood and inner material, which were collected aseptically and placed in sterile Ziplock plastic bags. These samples were then diluted in sterilized distilled water to attain 10-3 mg/ mL, and subsequently inoculated on Potato Dextrose Agar (PDA), Malt Extract Agar (MEA) and Czapek Dox Agar (CDA). The inoculants were incubated at 37 °C, followed by the observation of mold colony after the 11th day. The results identified seven and nine species of molds in the Kalisidi and Nglimut observation stations, respectively. The most abundant species was Penicillium sp. which was found in composted nest materials for the whole observation periods.

Pigment Produced by Glycine-Stimulated Macrophomina Phaseolina Is a (−)-Botryodiplodin Reaction Product and the Basis for an In-Culture Assay for (−)-Botryodiplodin Production

An isolate of Macrophomina phaseolina from muskmelons (Cucumis melo) was reported by Dunlap and Bruton to produce red pigment(s) in melons and in culture in the presence of added glycine, alanine, leucine, or asparagine in the medium, but not with some other amino acids and nitrogen-containing compounds. We explored the generality and mechanism of this pigment production response using pathogenic M. phaseolina isolates from soybean plants expressing symptoms of charcoal rot disease. A survey of 42 M. phaseolina isolates growing on Czapek-Dox agar medium supplemented with glycine confirmed pigment production by 71% of isolates at the optimal glycine concentration (10 g/L). Studies in this laboratory have demonstrated that some pathogenic isolates of M. phaseolina produce the mycotoxin (−)-botryodiplodin, which has been reported to react with amino acids, proteins, and other amines to produce red pigments. Time course studies showed a significant positive correlation between pigment and (−)-botryodiplodin production by selected M. phaseolina isolates with maximum production at seven to eight days. Pigments produced in agar culture medium supplemented with glycine, beta-alanine, or other amines exhibited similar UV-vis adsorption spectra as did pigments produced by (±)-botryodiplodin reacting in the same agar medium. In a separate study of 39 M. phaseolina isolates, red pigment production (OD520) on 10 g/L glycine-supplemented Czapek-Dox agar medium correlated significantly with (−)-botryodiplodin production (LC/MS analysis of culture filtrates) in parallel cultures on un-supplemented medium. These results support pigment production on glycine-supplemented agar medium as a simple and inexpensive in-culture method for detecting (−)-botryodiplodin production by M. phaseolina isolates.