MYCOLOGICAL ISOLATION AND IDENTIFICATION IN THE CAFETERIA OF HAWLER CITY KURDISTAN REGION, IRAQ

The availability and dynamics of airborne fungal spores in the atmosphere are strongly influenced by the meteorological parameters and by other factors such as air pollutants. The aim of this study was to update the knowledge about the seasonal and diurnal variations in airborne fungal spores of Doha area and to correlate these variations with meteorological factors, and to investigate the influence of atmospheric CO2 concentrations and different culture media on the availability of fungal spores. From 106 settle plate exposures (on alternative days) throughout the period April 2015-March 2016, a total of 1197 mould- and 283 yeast colony-forming units (CFU), twenty one genera and 62 species were retrieved. The highest fungal spore's concentration was recorded in February 2016, whereas the lowest concentration occurred in August 2015. The main constituents of the fungal airspora were attributed to Cladosporium (60.2%), Aspergillus (10.4%), Fusarium (9.4%), Alternaria (8.5%), and Ganoderma spp. (2.3%). Cladosporium showed two peaks in April and February, while Fusarium and Alteranria peaked in July. Aspirgillus had one peak in August. The prevalence of Ganoderma spp. were exclusively detected in February and March. Temperature was significantly and negatively correlated with the total colony count and fungal species, however no significant correlation was found between relative humidity and both the total colony count and fungal species. Wind speed was significantly and positively correlated with the total colony count and fungal species. The correlation between rainfalls and either total colony count or fungal species was non-significant. Intra-diurnal fluctuations of fungal spores was investigated during the period of 1st of Feb – 31st of March 2016. The highest dispersal of fungal spores favored 18:00 h, whereas at 00:00 h (midnight) the lowest fungal spores release was recorded. No significant difference was observed in total number of fungal colonies or species collected with the two media Potato Dextrose Agar (PDA) and Rose Bengal media. Nevertheless, certain fungal taxa were highly selective and thus their growth rate was on one media much higher than with another. There were no significant differences in the composition and diversity of the airborne fungal population between two different study sites under the influence of atmospheric CO2 concentration, though daily concentration of CO2 was higher at the Industrial area site than at Qatar University Campus. Remarkably, the concentrations of Alternaria spp. and Fusarium spp. were significantly higher at Industrial area site in corresponding to CO2 than at Qatar University site.

The present study was aimed to isolate and screen the ability of citric acid producing Aspergillus spp from soil. Citric acid is the most important organic acid, used as a natural preservative and conservative. It is also used to add an acidic or sour taste to foods and soft drinks. In this study, fungal samples were isolated from soil samples, using serial dilution agar plating method and the isolates were identified based on their microscopic and morphological characteristics. From the isolates, the dominant fungal species, Aspergillus niger and Aspergillus fumigatus were sub-cultured using Potato Dextrose Agar (PDA) medium and the fungal species were used for citric acid production. The spores were mixed in the fermentative medium for 72 hours at 30±1° C. After incubation, the citric acid content was estimated by titrimetric method. Among the two species, higher citric acid production was shown by Aspergillus niger S-6. Then it was undertaken for optimisation under various pH, temperature, Carbon and Nitrogen sources. In this assay, the maximum production of citric acid was observed in Aspergillus niger S-6 at pH 6.0, temperature 30±1° C, Carbon source Glucose and Nitrogen source Ammonium sulphate.

The purpose of this study was to separate microorganisms (fungi and bacteria) from three distinct vegetables: celery, lettuce, and arugula. The Potato Dextrose Agar (PDA) method of the power plate method was utilized to isolate and identify various fungal species, and the same procedure was followed to isolate bacteria from the samples under study. The findings identify the fungi and display the total number of colony forming units (CFU) in three different vegetables grown on PDA medium. Rhizopus sp., Aspergillus sp., Alternaria, yeast, Penicillium, Alternaria, and Rhodotorula were the most often isolated fungi. Mucor and Demaceous were the less common species of fungi found. The first grocery store had celery with a higher count of bacteria—268 CFU. With 250 CFU, lettuce had the highest amount of bacteria in the second grocery. Additionally, celery was found to have a higher concentration of bacteria in the third grocery store—280 CFU.

Fungal-feeding habits of six nematode isolates in the genus Filenchus

Cannabis products are subjected to microbial testing for pathogenic fungi and bacteria. These testing requirements often rely on non-specific colony forming unit (CFU/g) specifications without clarity on which medium, selection or growth times are required. We performed whole genome sequencing to assess the specificity of colony forming units (CFU) derived from three different plating media: Potato Dextrose Agar (PDA), PDA with chloramphenicol and Dichloran Rose Bengal with chloramphenicol (DRBC). Colonies were isolated from each medium type and their whole genomes sequenced to identify the diversity of microbes present on each medium selection. Fungal Internal Transcribed Spacer (ITS3) and Bacterial 16S RNA(16S) quantitative polymerase chain reactions (qPCR) were performed, to correlate these CFUs with fungi- and bacterial- specific qPCR. Each plating medium displayed a ten-fold difference in CFU counts. PDA with chloramphenicol showed the highest diversity and the highest concordance with whole genome sequencing. According to ITS3 and 16S qPCR confirmed with whole genome sequencing, DRBC under counted yeast and mold while PDA without chloramphenicol over counted CFUs due to bacterial growth without selection. Colony Forming Unit regulations lack specificity. Each medium produces significant differences in CFU counts. These are further dependent on subjective interpretation, failure to culture most microbes, and poor selection between bacteria and fungi. Given the most pathogenic microbes found on cannabis are endophytes which culture fails to detect, molecular methods offer a solution to this long-standing quantification problem in the cannabis testing field.

Fungal diversity in the seawater and sediment in the mangrove forest at Andaman Coastal Research Station for Development, Ranong Province was studied. Seawater and sediment were sampled in 5 sites during the rainy and dry season. The fungi were cultivated in Potato Dextrose Agar (PDA) supplemented with Streptomycin and Chloramphenical and incubated at room temperature for 7 days. Eighty nine and 99 fungal isolates were found in the sediment and seawater samples, respectively. Fungal identification was done using their molecular study. Forty-five fungal species could be identified using molecular study and morphological features including: Acremonium furcatum, A. nepalense, Aspergillus aculeatus, A. flavus, A. fumigatus, A. niger, A. nomius, A. nutans, A. oryzae, A. tubingensis, Aspergillus sp., Ceratocystis paradoxa, Cladosporium cladosporioides, C. oxysporum, Colletotrichum gloeosporioides, Diaporthe helianthi, Didymellaceae sp., Dothideomycetes sp., Eupenicillium shearii, Fusarium equiseti, F. moniliformis, F. nelsonii, F. oxysporum, F. proliferatum, F. solani, F. subglutinans, Fusarium sp., Ganoderma cupreum, Gliocephalotrichum simplex, Gongronella butleri, Hypocrea jecorina, Lasiodiplodia theobromae, Neosartorya fischeri, Penicillium citrinum, P. funiculosum, P. griseofulvum, P. janthinellum, P. lilacinum, P. oxalicum, P. rademirici, P. simplicissimum, P. stecki, P. vasconiae, Penicillium sp., Pestalotiopsis mangiferae, Pleurotus pulmonarius, Rigidoporus vinctus, Sordaria sp., Talaromyces assiutensis, Trichaptum laricinum, Trichoderma asperellum, Trichoderma sp. and Xylaria apoda. The common fungal species found in this study were Penicillium, Aspergillus and Fusarium.

The fungi use the various substrates such as carbon and energy source, are one of the major causative agents biodeterioration of bibliographic materials in archives and libraries. In the Universidad del Valle, a study of fungal populations with the aim of isolating the fungi on the surface of the books and in the environment of the library. We performed a sampling of books with signs of bio-deterioration of four sections (General Books Store, Antiques Books Collection, Periodical Library Collection and Periodical Library Store). A total of 409 colony-forming units (UFC) on potato dextrose agar and cellulose agar were isolated, locating (89.7%) in the environment and in the books (10.3%). The 37.16% of UFC were detected in the General Books Store, 9.78% at Periodical Library and a 34.97% at Antiques Books Collection. There were 17 genera of fungi as the most abundant, Cladosporium (59.72%), Fusarium (9.31%), Curvularia (6.62%), Aspergillus (6.37%) and Chaetomium (5.64%). All genera showed ability to grow in agar cellulose. The Kruskal-Wallis test (p=0.552) determined that there were no significant differences in the number of colonies in UFC/m2/min isolated environment for the four sections of the Library.

Aim: To carry out a Comparative mould analysis using groundnut shell infusion agar (GSA) and potato dextrose agar (PDA), as the control.
 Study Design: Laboratory-experimental design was used in this study.
 Place and Duration of Study: Soil samples were obtained from three different locations (Garden soil beside Biology Main Laboratory, opposite Faculty of Law and Faculty of Agriculture) in Rivers State University, Port Harcourt Rivers State, Nigeria. The study was carried out for three (3) months at the Microbiology Laboratory, Rivers State University, Port Harcourt.
 Methodology: Groundnut Shell Infusion Agar (GSA) was prepared by weighing 28 g of blended gari and 15 g of agar powder into 1L of groundnut shells filtrate. Potato dextrose Agar (PDA), a conventional medium was prepared according to the manufacturer’s specifications. GSA was prepared by weighing 28 g of blended gari and 15 g of agar powder into 1L of groundnut shells filtrate. Potato dextrose agar, a conventional medium was prepared according to the Manufacturer’s specifications.
 Results: The mean mould counts from the different locations ranged from 3.7×107 cfu/ml to 7.8×107 cfu/ml on GSA and 3.7×107 cfu/ml to 1.5×109 cfu/ml on PDA following incubation at room temperature (27°c ± 2) for 3-5 days. The moulds identified were Aspergillus niger, Aspergillus flavus, Trichoderma viride, Rhizopus sp. Mucor sp. Botrytis sp. Helminthosporium caryopsidum, and Penicillum sp.
 Conclusion: From the results obtained, it showed that GSA could be used successfully for quantitative mould counts and other mycological studies. This would proffer solution to the high cost of conventional media used for moulds as well as agro waste pollution in the environment.

This is the first study to isolate and identify the aeromycoflora of Jizan province . The study was carried out to estimate quantitatively and qualitatively the mycoflora present in the air of different sampling sites of the province. Open petri plate method with Potato Dextrose agar media was used with an exposure time of 2 mins at each of the seven sampling sites. Total number of eight genera were isolated from the seven indoor and outdoor sampling sites of the province represented by Aspergillus niger, Aspergillus sp. , Chaetomium ,Cladosporium, Nigrospora,Penicillium , Rhodotorula , Scedosporium and Ulocladium. Seventy two fungal isolates were obtained from the eight represented genera . Aspergillus was the predominant genera.The Colony forming unit (CFU) 3 was calculated for all of the sampling sites under study to know the level of contamination .The high wind speed and the busy hours of business were the major factors responsible for large number of fungal counts during this study from the province.

Background: Cannabis products are subjected to microbial testing for human pathogenic fungi and bacteria. These testing requirements often rely on non-specific colony forming unit (CFU/g) specifications without clarity on which medium, selection or growth times are required. We performed whole genome sequencing to assess the specificity of colony forming units (CFU) derived from three different plating media: Potato Dextrose Agar (PDA), PDA with chloramphenicol and Dichloran Rose Bengal with chloramphenicol (DRBC). Methods: Colonies were isolated from each medium type and their whole genomes sequenced to identify the diversity of microbes present on each medium selection. Fungal Internal Transcribed Spacer (ITS3) and Bacterial 16S RNA(16S) quantitative polymerase chain reactions (qPCR) were performed, to correlate these CFUs with fungi- and bacterial- specific qPCR. Results: Each plating medium displayed a ten-fold difference in CFU counts. PDA with chloramphenicol showed the highest diversity and the highest concordance with whole genome sequencing. According to ITS3 and 16S qPCR confirmed with whole genome sequencing, DRBC under counted yeast and mold while PDA without chloramphenicol over counted CFUs due to bacterial growth without selection. Conclusions: Colony Forming Unit regulations lack specificity. Each medium produces significant differences in CFU counts. These are further dependent on subjective interpretation, failure to culture most microbes, and poor selection between bacteria and fungi. Given the most human pathogenic microbes found on cannabis are endophytes which culture fails to detect, molecular methods offer a solution to this long-standing quantification problem in the cannabis testing field.

Cannabis products are subjected to microbial testing for human pathogenic fungi and bacteria. These testing requirements often rely on non-specific colony forming unit (CFU/g) specifications without clarity on which medium, selection or growth times are required. We performed whole genome sequencing to assess the specificity of colony forming units (CFU) derived from three different plating media: Potato Dextrose Agar (PDA), PDA with chloramphenicol and Dichloran Rose Bengal with chloramphenicol (DRBC). Colonies were isolated from each medium type and their whole genomes sequenced to identify the diversity of microbes present on each medium selection. Fungal Internal Transcribed Spacer (ITS3) and Bacterial 16S RNA(16S) quantitative polymerase chain reactions (qPCR) were performed, to correlate these CFUs with fungi- and bacterial- specific qPCR. Each plating medium displayed a ten-fold difference in CFU counts. PDA with chloramphenicol showed the highest diversity and the highest concordance with whole genome sequencing. According to ITS3 and 16S qPCR confirmed with whole genome sequencing, DRBC under counted yeast and mold while PDA without chloramphenicol over counted CFUs due to bacterial growth without selection. Colony Forming Unit regulations lack specificity. Each medium produces significant differences in CFU counts. These are further dependent on subjective interpretation, failure to culture most microbes, and poor selection between bacteria and fungi. Given the most human pathogenic microbes found on cannabis are endophytes which culture fails to detect, molecular methods offer a solution to this long-standing quantification problem in the cannabis testing field.

Abstract. Nofiani R, Mu’in R, Hafizah, Ardiningsih P. 2024. Bioremoval of Pb2+ by Aspergillus niger D1RA, A heavy metal-resistant fungus isolated from an illegal gold mining site. Biodiversitas 25: 2504-2511. Heavy metal pollution can cause serious problems for the environment and human health. One of the methods to eliminate this pollution is to use heavy metal-resistant fungi isolated from heavy metal-polluted environments. This study aimed to investigate the ability of heavy metal-resistant fungi to remove Pb2+ in liquid media, Potato Dextrose Broth (PDB). Samples were collected from two locations, namely an abandoned illegal gold mining site and illegal gold mine, Samalantan, Bengkayang District, West Kalimantan, Indonesia. Each sample was inoculated on two different agar media (PDA= Potato Dextrose Agar and MEA= Malt Extract Agar) supplemented with 7.5 ppm HgCl2. All fungal species that grew on the surface media were isolated, identified (based on spore morphology and Internal Transcribed Spacer [ITS]), evaluated (tolerance index [TI] against Hg2+, Pb2+, and Zn2+), and assessed for their bioaccumulation capacity and Pb2+removal efficiency. Four isolates (Aspergillus sp. OK2A, Aspergillus sp. OEA, Aspergillus sp. OEB and OEC) were successfully isolated from the abandoned illegal gold mine, while only one isolate (Aspergillus niger D1RA) was isolated from the illegal gold mining site. On the eighth day of incubation, the high tolerance level of each fungus to various selected metal concentrations was Aspergillus sp. OK2A in 40 ppm HgCl2 and 300 ppm ZnCl2; Aspergillus sp. OEA in 40 ppm HgCl2 and 1,200 ppm ZnCl2; Aspergillus sp. OEB in 800 ppm Pb(NO3)2; OEC in 20 ppm HgCl2; A. niger D1RA in 40 ppm HgCl2, 1,200 ppm Pb(NO3)2, 300 ppm ZnCl2. Only A. niger D1RA showed a high tolerance for three metals and was further analyzed to determine the bioaccumulation capacity and removal efficiency of Pb2+. The best bioaccumulation capacity and removal efficiency of Pb2+ in PDB medium supplemented with 100 ppm Pb(NO3)2 at pH 4 were 237.776 mg/g dried biomass and 93.266 %, respectively. In conclusion, A. niger D1RA has the potential as a bioremediation agent to remediate Pb2+ environments.

Anona muricata commonly known as sour-sop, belong to the family Annonaceae. It is a small slender tropical tree usually grown for its large fleshy and juicy fruits. The fruit of A. muricata plays an important role in the diet of many in many parts of the tropics including Nigerian. Unfortunately, the usefulness of the fruits of Anona is decimated by many fungi species. Investigation of the fungi that cause the deterioration of sour-sop (Anona muricata) was carried out in order to recommend the appropriate control measures. Mature fruits of A. muricata were collected from different locations in Abia State. Isolation, characterization and identification of fungi were made by plating washings from skin surface and extracted juice from pulp of the fruits in test tubes containing potato dextrose agar (PDA) and yeast malt extract agar (Difco) into which streptomycin sulphate was incorporated to inhibit bacterial growth. Inoculated tubes were incubated at 28 ± 2°C for 48 h. Control experiment was carried out with sterile peptone water instead of using the washings. The following filamentous and yeast fungi were isolated from the skin surface and pulp of the fruits: Aspergillus flavus, Aspergillus niger, Botryodiplodia theobromae, Colletotrichum sp., Fusarium solani, Mucor sp., Penicillium chrysogenium, Penicillium sp., Rhizopus stolonifer and Rigidoporus sp., Candida albicans, Saccharomyces cerevisiae, Saccharomyces rouxii and Torulopsis spp. The pathogenicity tests of the filamentous fungi isolated were carried out on mature green Anona fruits and were found to be pathogenic. The filamentous fungi were mainly responsible for the deterioration of the fruits of A. muricata in Abia state while the yeasts were fermentative. Key words: Abia state, Anona muricata (Sour-sop) fruits, fungi, deterioration.

Grapes kept after harvest for withering in drying fruit rooms (fruttaio) are highly susceptible to Botrytis cinerea infection. A taxonomically undefined isolate (B83) was identified during a survey of B. cinerea populations in these specific environments. This isolate yielded negative results in B. cinerea‐specific PCRs and displayed a different colony morphology on potato dextrose agar. Based on this data, the isolate underwent a taxonomic investigation. Sequencing of the ITS–5·8S–ITS2 region indicated that the isolate belongs to the Botrytis genus. IGS‐RFLP analysis revealed that Botrytis sp. B83 was a different haplotype from genotypes of indigenous B. cinerea strains. Comparative sequence analysis of three housekeeping genes, G3PDH, HSP60 and RPB2, showed that the isolate constitutes a taxon related to Botrytis aclada, distant from B. cinerea. A phylogenetic tree built with NEP genes was in agreement with the combined tree built with G3PDH, HSP60 and RPB2 sequences. Botrytis sp. B83 is morphologically similar to B. aclada. Conversely, its host range indicates that it is polyphagous like B. cinerea, despite its lower virulence. This study provides evidence for the existence of a new species of Botrytis. Nevertheless, the description of only a single isolate and the need for further information on genetic divergence, reproductive isolation and host preference, prevent the classification and formal description of a new taxon.

During May 2016, a corky textured, "star like" symptom, located at the apex on the far side of the fruit was observed on young persimmon fruitlets (Fig. 1), on several persimmon varieties in plantations located along the Mediterranean Sea coast. The lesions caused cosmetic damage, which disqualified the fruit from marketing and can affect as much as 50% of the fruit in the orchard. Symptoms were correlated with the presence of wilting flower parts (petals and stamens) attached to the fruitlet (Fig. 1). Fruitlets with no flower parts attached did not develop the corky star symptom, while almost all fruitlets with an attached wilted flower parts had symptoms underneath the flower parts. Flower parts and fruitlets displaying the phenomenon were sampled (orchard near the town of Zichron Yaccov) and used for fungi isolation. At least ten fruitlets were surface sterilized by immersion in 1% NaOCl for 1 min. Pieces of the infected tissue were then placed on 0.25% potato dextrose agar (PDA) supplemented with 12 µg/mL tetracycline (Sigma, Rehovot, Israel). In addition, the inner parts of at least ten moldy flowers were placed on 0.25% PDA supplemented with tetracycline and incubated at 25°C for 7 days. Two fungi were isolated from the flower parts and the symptomatic fruitlets: Alternaria sp. and Botrytis sp. Koch postulates were conducted by inoculating 10 µl of conidial suspension of each fungus (105 conidia/ml in H2O, single spore originated), on four wounds, made by puncturing a 2 mm deep hole with 21G sterile syringe needle, on the apex of surface sterilized, small, green fruits. Fruits were placed in sealed 2-liter plastic boxes. Symptoms similar to those found on the fruitlets in the orchards were observed on the fruit inoculated with Botrytis sp. (corky like, but not in the shape of stars) about 14 days post inoculation. Botrytis sp. was re-isolated from the symptomatic fruit to fulfill Koch's postulates. Alternaria and water inoculation did not cause any symptoms. The Botrytis sp. colonies, when grown on PDA, grow initially as white colonies becoming gray to brown after about seven days. Elliptical conidia, 8 to 12 μm long and 6 to 10 μm wide, were observed under light microscope. Isolate Pers-1, incubated at 21°C for 21 days, produced blackish, spherical to irregular microsclerotia, ranging from 0.55 to 4 mm (width and length, respectively). For molecular characterization of the Botrytis sp. isolate, Pers-1, fungal genomic DNA was extracted as previously described by Freeman et al. (2013). The internal transcribed spacer (ITS) sequence region of rDNA was amplified using ITS1/ITS4 primers (White et al. 1990), and sequenced. ITS analysis revealed that it belongs to genus Botrytis (MT573470.1 with 99.80% identity). For further confirmation, nuclear protein-coding genes (RPB2 and BT-1, Malkuset et al. 2006 and Glass et al. 1995) were sequenced and found to have 99.87% and 99.80% identity to Botrytis cinerea Pers. Sequences, deposited in GenBank as accessions OQ286390, OQ587946 and OQ409867, respectively. Botrytis was previously reported to cause persimmon fruit scarring and damage of calyces (Rheinländer et al. 2013) and fruit rot during post-harvest (Barkai-Golan. 2001), yet to the best of our knowledge, this is the first report of B. cinerea causing "star like" corky symptoms on persimmon in Israel.