Cladosporium Spp Research Articles

Gray mold disease on bottle gourd caused by Cladosporium tenuissimum in China.

Bottle gourd [Lagenaria siceraria (Mol.) Stand] is a widely cultivated succulent crop species. In December 2022, a serious bottle gourd disease occurred in the protected vegetable planting base of Xingguo County, Ganzhou City, Jiangxi Province, China, with 85% of the 2,100 plants having gray mold disease-like symptoms, including gray spots on the infected fruit. They quickly expanded at suitable temperature and humidity, forming a gray mold layer with inward depressions, which spread to the fruit stem causing watery rot, and the flesh turned black and started to rot. To isolate the pathogen, fruits of the diseased plants were surface-disinfected with 75% ethanol for 30 s, immersed in 0.1% HgCl2 for 1min, rinsed thrice with sterile water, and cultured on a potato-dextrose agar (PDA) medium at 28°C. Mycelia from the diseased tissue were subcultured on fresh PDA medium to obtain pure cultures. After incubation at 25°C for 7 days, olive-green colonies (~2.5mm·d-1) developed. Cultures developed numerous elliptical and limoniform conidia measuring 2.69~9.79 μm to 2.10~5.92 μm (average 5.62×3.12 μm) (n=20). The morphological characteristics of the pathogen resembled those of Cladosporium spp. Fungal genomic DNA was extracted, and the internal transcribed spacer (ITS), partial translation elongation factor-1 alpha (TEF-1α), and actin (ACT) regions were amplified with primers ITS1/4, TEF-728F/986R, and ACT-512F/783R, respectively, and sequenced (Bensch et al. 2012; Jo et al. 2018). Basic Local Alignment Search Tool analysis (BLAST) revealed that the ITS (accession no. OQ186729), ACT (OQ240962), and TEF-1α (OQ240963) sequences of isolate hjt4 shared the highest similarity (99-100%) with those of Cladosporium tenuissimum (accessions no. OM232068, OM256530, OM256526) (Duccio et al. 2015). A phylogenetic tree of the isolate hjt4 and its close relatives within Cladosporium was constructed using the MEGA X neighbor-joining method. The pathogen was identified as C. tenuissimum based on morphological and molecular characteristics. A specimen (JXAU-H2022982) was deposited at the Herbarium of the College of Agronomy, Jiangxi Agricultural University. To confirm its pathogenicity, seven-day-old healthy bottle gourd fruits were disinfected with 75% ethanol, 1 mm-deep wounds were made with sterilized scalpels, and the plants were inoculated with PDA plugs (0.8 cm in diameter) containing actively growing mycelia of isolate hjt4. Plants inoculated with sterile PDA plugs served as controls. Each group contained three fruits, and the experiment was performed in triplicate. All fruits were incubated in a biochemical incubator at 28°C. After 3 days, the fruit surface shrank, and the flesh turned to a black colour and rotten, which rapidly spread to the branches. Control fruits did not develop any symptoms. Reisolated colonies showed the same morphological traits as those of the inoculation isolates, whereas no target colonies were isolated from the control fruits. The pathogen was previously reported to cause leaf blight disease in Coriandrum sativum (Zhou et al. 2022) and sooty spots on Cape gooseberry (Miyake et al. 2022), among others. To our knowledge, this is the first report of gray mold disease caused by C. tenuissimum on bottle gourd in China. The findings provide an important foundation for monitoring and controlling the spread of this disease.

Study of the Microbiome of the Cretan Sour Cream Staka Using Amplicon Sequencing and Shotgun Metagenomics and Isolation of Novel Strains with an Important Antimicrobial Potential.

Staka is a traditional Greek sour cream made mostly from spontaneously fermented sheep milk or a mixture of sheep and goat milk. At the industrial scale, cream separators and starter cultures may also be used. Staka is sometimes cooked with flour to absorb most of the fat. In this study, we employed culture-based techniques, amplicon sequencing, and shotgun metagenomics to analyze the Staka microbiome for the first time. The samples were dominated by Lactococcus or Leuconostoc spp. Most other bacteria were lactic acid bacteria (LAB) from the Streptococcus and Enterococcus genera or Gram-negative bacteria from the Buttiauxella, Pseudomonas, Enterobacter, Escherichia-Shigella, and Hafnia genera. Debaryomyces, Kluyveromyces, or Alternaria were the most prevalent genera in the samples, followed by other yeasts and molds like Saccharomyces, Penicillium, Aspergillus, Stemphylium, Coniospotium, or Cladosporium spp. Shotgun metagenomics allowed the species-level identification of Lactococcus lactis, Lactococcus raffinolactis, Streptococcus thermophilus, Streptococcus gallolyticus, Escherichia coli, Hafnia alvei, Streptococcus parauberis, and Enterococcus durans. Binning of assembled shotgun reads followed by recruitment plot analysis of single reads could determine near-complete metagenome assembled genomes (MAGs). Culture-dependent and culture-independent analyses were in overall agreement with some distinct differences. For example, lactococci could not be isolated, presumably because they had entered a viable but not culturable (VBNC) state or because they were dead. Finally, several LAB, Hafnia paralvei, and Pseudomonas spp. isolates exhibited antimicrobial activities against oral or other pathogenic streptococci, and certain spoilage and pathogenic bacteria establishing their potential role in food bio-protection or new biomedical applications. Our study may pave the way for additional studies concerning artisanal sour creams to better understand the factors affecting their production and the quality.

Mycological and Mycotoxicological Producing Potential of Isolates from Fermented Melon Seeds [Citrullus lanatus (Thumb) Matsun ‘Egusi kirikiri’

This study identified and examined fungal presumed producing mycotoxins in fermented melon seed condiment (ogiri). The samples were cultured on Potato Dextrose Agar and Sabouraud Dextrose agar to identify fungi. The detection of mycotoxins in the fermented melon seeds samples was carried out by the use of Coconut Agar Media (CAM), by which the fungal isolates were cultured and plates incubated for 3-7days at 280C. The mycobiota of fermented melon seed revealed of many species belonging to the Aspergillus spp, Penicillium spp, Cladosporium spp, and Rhizopus oryzae. Among all the fungi identified in the study, Aspergillus spp has the highest prevalence rate (44.4%) while Penicillium spp, and Rhizopus oryzae have same value (22.2%).the lowest is Cladosporium spp with (11.1%) and its presence can be attributed to decaying of the samples. The results obtained show that some of the fungal isolates had the ability to produce mycotoxins. From the mycotoxin screening, only six (6) species (Aspergillus spp, and Penicillium spp) were positive (+), Cladosporium spp, and Rhizopus oryzae were negative (-). Mycotoxin contamination level varied in different market locations in which Swali Market had the highest level of mycotoxin (50%) followed by Opolo Market (33.3%) and Tombia Market (16.7%). Mycotoxigenic producing fungi and mycotoxin levels in fermented melon seed samples are public health concern. Meanwhile, the occurrence of the pathogenic fungi (Aspergillus, Penicillium, and Rhizopus) in fermented melon seed can affect its shelf life and can also cause severe health challenges.

On the Way to the Technological Development of Newly Selected Non-Saccharomyces Yeasts Selected as Innovative Biocontrol Agents in Table Grapes.

Post-harvest decay of fresh table grapes causes considerable annual production losses. The main fungal agents of decay both in pre- and post-harvest are B. cinerea, Penicillium spp., Aspergillus spp., Alternaria spp., and Cladosporium spp. To date, the use of agrochemicals and SO2 are the main methods to control grape molds in pre- and postharvest, respectively. Significant improvements, however, have already been made in to apply innovative and more environmentally sustainable control strategies, such as Biological Control Agents (BCAs), which can reduce disease severity in both pre- and post-harvest. In this study, 31 new non-Saccharomyces yeast strains, isolated from berries of native Apulian table grape genotypes, were tested for their in vivo effectiveness against grey mold of table grapes, resulting in two St. bacillaris ('N22_I1' and 'S13_I3'), one S. diversa ('N22_I3'), one A. pullulans ('OLB_9.1_VL') and one H. uvarum ('OLB_9.1_BR') yeast strains that were marked as efficient and good BCAs. Their mechanisms of action were characterized through in vitro assays, and additional characteristics were evaluated to assess the economic feasibility and viability for future technological employment. Their effectiveness was tested by reducing the working concentration, their antagonistic effect on a wide range of fungal pathogens, their ability to survive in formulations with long shelf life, and their safety to human health.

Identification and Characterization of Fungal Pathogens Contributing to Citrus Twig Dieback in Kerman Province, Iran

Citrus decline with complex symptoms was recently observed in Kerman province, reducing the annual yield of citrus fruits. Agents that cause citrus decline include fungi, bacteria, viruses, nematodes, and abiotic factors. One of the most important symptoms of citrus decline is branch dieback. In this study, the fungal agent responsible for dieback was identified. To achieve this goal, sampling was conducted in citrus orchards with symptoms of twig dieback and twig canker in the summer of 2020. Samples were collected from different areas of the citrus growing region of Kerman province, including Bam, Rigan, Narmashir, Fahraj, Shahdad, and Orzueeh. Fungi were isolated from diseased branches on nutrient culture medium. The resulting fungi were identified using morphological characteristics. The following fungi were isolated from diseased plants: Alternaria spp., Nattrassia mangiferae, Ulocladium spp., Bipolaris spp., Fusarium spp., Phoma sp., Rhizoctonia sp., Paecilomyces spp., Chaetomium spp., Acremonium spp., Stemphylium sp., and Cladosporium spp. Notably, Alternaria spp. exhibited the highest frequency and widest distribution among them, making it the most prevalent fungus throughout all citrus-growing regions of Kerman province. N. mangiferae was the second most frequently isolated fungus. Pathogenicity tests were performed by artificially inoculating two-year-old lemon trees under controlled conditions. All fungal genera except Chaetomium sp. and Cladosporium spp. were virulent. N. mangiferae was the most virulent fungus and caused the most severe cankers six weeks after inoculation. It also had the greatest discoloration compared to other fungi in these tests

Efficiency of Some Clays and Organic Materials on the Reduction of Aflatoxin B1 Produced from Isolates of the Fungus Aspergillus flavus Contaminating Corn Grains

The study was conducted to evaluate the effectiveness of certain types of clay minerals, activated charcoal made from common reed and date palm kernel, in reducing the levels of aflatoxin B1(AFB1) produced by Aspergillus flavus that contaminated corn grains in a liquid medium, Yeast Extract Sucrose (YES). The results showed the presence of nine species of fungal genera that contaminated corn grains, with the Aspergillus spp being the most predominant, accounting for 42%, followed by the Penicillium spp with 27%, and the Fusarium spp with a 21%. The genera Rhizopus spp, Mucor spp, and Cladosporium spp had a prevalence of 5%, 4%, and 1%, respectively. The results of the thin-layer chromatography (TLC) test, which was used to isolate A. flavus fungi from corn grains and detect the presence of AFB1, showed variations in their ability to produce AFB1. Seventy-five percent of the diagnosed A. flavus isolates showed the ability to produce AFB1. The results indicated that all types of minerals used in the study were effective in reducing the levels of AFB1 in liquid media. Top of Form Similarly, activated charcoal from common reed showed a high potential for AFB1 reduction, where the highest reduction percentage was 93.90% for the activated charcoal from fermented reed at a concentration of 30 mgml−1.

Assessment of Microbial Air Quality of Antenatal and Postnatal Wards of Four Major Hospitals in Ondo State Nigeria

The air microbial load and antibiotic susceptibility of microorganisms obtained from antenatal and postnatal wards of four major hospitals in Ondo State, Nigeria were assessed. The microbial load of each ward was determined by the passive settle plate method. The bacterial and fungal isolates obtained were identified based on cultural, morphological, and biochemical characteristics. The following microorganisms were tentatively identified: Proteus spp, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp, Micrococcus luteus, Pseudomonas spp, Bacillus subtillis, Bacillus cereus, Aspergillus spp, Penicillium notatum, Cladosporium spp, Mucor spp and Articulospora inflata. The bacteria that had the highest percentage occurrence (28.50%) were Bacillus subtillis and Staphylococcus aureus, while the least occurrence was recorded for Serratia marcescens (2.38%). Among the fungi, Penicillium notatum had the highest occurrence (44.70%), while Articulospora inflata (1.18%) had the lowest occurrence. The result of the antibiotic sensitivity pattern of the isolates showed that all the bacteria were susceptible to the commercial antibiotics except Serratia marcescens which were resistant to all the tested antibiotics. It is well known that life-threatening infections can be acquired from the hospitals, hence, it is recommended that improved hygienic procedures be put in place in these hospitals to reduce the occurrence of these microorganisms in antenatal and postnatal wards.

Green Biocidal Nanotechnology Use for Urban Stone-Built Heritage—Case Study from Oradea, Romania

Heritage buildings clad with natural rock endure over time destruction caused by weathering mechanisms, pollution from urban areas, biodeterioration due to organisms, microorganisms, and also the anthropic factor. On the surface of the limestone samples taken from the ornamental natural rock with which the outside of Markovits-Mathéser house, Oradea, Romania, is clad, two species of fungi were inoculated in the laboratory: Aspergillus spp. and Cladosporium spp. Wollastonite was then applied, and from the imaging analysis (SEM), the inhibition of fungi by it is clearly observed (48 h after its application), which was also confirmed using the image segmentation method. It was also noted that the hydrophilicity of the aqueous suspension of wollastonite resulted in the absorption of water in the substrate, which in turn resulted in the drying out and surface breakage of the specimens. X-ray diffraction analysis showed the presence of the two phases (calcite and quartz) as in the starting sample, and also an additional phase assigned to wollastonite in the later phase of the experiment. An amorphous component, due to the applied gel composition, was also reported. This research highlights the fact that there are good premises for aqueous suspension of wollastonite to have a biocidal character for Aspegillus spp. and Cladosporium spp., when applied on natural stone used in the construction of heritage buildings located in temperate climates; due to its easy application, green and ecofriendly properties, and also low cost of acquisition and application.

Quantifying the mycobiome and its major constituents on the skin of 20 normal dogs.

To report the density, and constituents, of the mycobiome on the skin surface of normal dogs. 20 normal dogs were recruited for this study, with informed consent in all cases. Flocked swabs were used to sample the skin surface and to sample the skin surface after superficial scraping with a blunted scapula. Both samples were taken within a brass guide with an internal area of 3.5 cm-2. Next-generation DNA sequencing was used to identify and quantify components of the mycobiome. The median density of the mycobiome was 1.1 X 105 cm-2 (IQR, 27,561, 409,572). Cladosporium spp and Vishniacozyma victoriae were found on all 20 dogs. Knowledge of the density and the composition of the cutaneous mycobiome will increase our understanding of skin biology and may have relevance to future therapeutic trials.

Influence of pH, agitation speed, and temperature on growth of fungi isolated from Koya, Iraq

Fungi are intensely affected by physical and physicochemical factors that affect their growth rate. The present study investigates the influence of different pH values, agitation speeds, and temperature ranges (ºC) on fungi growth rates by measuring their fresh and dry weight. Several fungi were isolated from different sites of Koya city, Iraq. After being cultured in a potato dextrose broth (PDB) medium, their measurements were taken. The results of the study showed growth of isolated fungi in all factors. Furthermore, the maximum growth is determined at pH 6, agitation speed of 70 ​rpm, and 25 ​°C. The p-value was calculated based on the correlation between the “fresh” and “dry” values for each of the fungi based on the effect of varying pH levels, agitation speeds, and temperatures (p ​= ​0.028277, 0.00065, and 0.016806, respectively). The statistical analysis revealed that there was statistically significant (p ​> ​0.05) difference in fungi growth rates based on the variations in pH, agitation speeds, and temperature. The study concluded that different growth circumstances had an impact on biomass growth in different species of fungi and optimal growth was shown by Fusarium begonia, Mucor spp, and Cladosporium spp.

Surveying potentially antagonistic fungi to myrtle rust (Austropuccinia psidii) in Brazil: fungicolous Cladosporium spp.

The myrtle rust (MR), caused by Austropuccinia psidii, is a worldwide threat to the cultivated and wild Myrtaceae. Originally from the neotropics, it has spread to North America, Africa, and Asia and has reached geographically isolated areas in the Pacific and Australasia. It is attacking native species in those new ranges and is still spreading and causing great concern for the damage caused to endemic Myrtaceae, and to the environment. Classical biological control is regarded as the most sustainable management option for mitigating such biological invasions. However, there are no examples of introductions of host-specific co-evolved natural enemies of plant pathogens, from their native range, as a management strategy for plant pathogens. In order to explore this neglected approach, a survey of potential fungal natural enemies of A. psidii was initiated recently in the state of Minas Gerais (Brazil). Several purported mycoparasites have been collected from A. Psidii pustules formed on myrtaceous hosts. This included some isolates of dematiaceous fungi recognized as having a Cladosporium-like morphology. Here we present the results of the investigation aimed at elucidating their identity through a polyphasic taxonomic approach. Besides morphological and cultural features, molecular analyses using sequences of translation elongation factor 1-α (EF1) and actin (ACT) were performed. The combination of data generated is presented herein and placed all Cladosporium-like isolates in six species of Cladosporium, namely, Cladosporium angulosum, C. anthropophilum, C. bambusicola, C. benschii, C. guizhouense, and C. macadamiae. None of these have ever been recorded in association with A. psidii. Now, with the identification of these isolates at hand, an evaluation of biocontrol potential of these fungi will be initiated. In contrast with the ready finding of fungicolous (possibly mycoparasitic) fungi on MR in this study, no evidence of those was recorded from Australasia until now.

Bruchus rufimanus Boh. Effect on Broad Bean Seed Quality and the Infection Level of Seed-Borne Fungal Pathogens.

Broad bean weevil (Bruchus rufimanus Boh.) is considered an economically important insect pest of the broad bean crop. Its damage starts when larvae burrow into the seed and feed on endosperm or kill the embryo, thus the seed cannot germinate. The microbiological quality and consumer safety of broad bean may be compromised by holes. This study was performed during 2018-2020 to estimate the effect of seed damage by B. rufimanus on seed quality, germination rate and seedling growth, and on the incidence of seed-borne fungal pathogens. Broad bean seeds were collected and classified as damaged by B. rufimanus or non-damaged. There was a relationship between broad bean weevil damage and an increase in fungal contamination. Results showed that germination decreased from 97.2% for non-damaged seeds to 81.4% for the seeds with B. rufimanus damage. Seven fungal genera were identified: Fusarium spp., Alternaria spp., Aspergillus spp., Penicillium spp., Cladosporium spp., Mucor spp. and Botrytis spp. The most common fungal genus was Cladosporium spp. Our research shows that B. rufimanus negatively affects the germination of its host's seeds and decreases the development of seedlings.

Cutaneous phaeohyphomycosis in a greater bulldog bat (Noctilio leporinus) in northeastern Brazil

An adult male greater bulldog bat (Noctilio leporinus) was found dead in a suburban area in the municipality of Patos, Paraiba, northeastern Brazil. At post-mortem examination, the bat was emaciated and had multifocal to coalescent grey, crusted, dry, scaly cutaneous lesions, irregularly distributed over the dorsal thoracoabdominal region, muzzle, labial commissures, ears and dorsoventral surfaces of the patagia. Histopathology revealed numerous longitudinal and transverse sections of fungal organisms, with weakly basophilic walls, associated with multifocal areas of ulceration of the epidermis, necrosis, rupture and discontinuity of collagen fibres in the dermis without any inflammatory response. Molecular identification matched the organism to Cladosporium spp, Curvularia spp, Exserohilum spp, Bipolaris spp (100%) and Alternaria spp (97%), all of which have been associated with phaeohyphomycosis. Phaeohyphomycosis should be included as a differential diagnosis of cutaneous lesions in chiropterans.

Novel N-acylsulfonamides: Synthesis, in silico prediction, molecular docking dynamic simulation, antimicrobial and anti-inflammatory activities

Microbial resistance to drugs currently traded in the market is a serious problem in modern medicine. In this field of research, we synthesized a novel N-acylsulfonamides (NAS) derivatives starting from commercially available compounds; morpholine, isocyanate of chlorosulfonyl and alcohols. The in vitro antimicrobial potential of synthesized compounds was screened against 04 Gram-negative bacteria; Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, 02 Gram-positive bacteria: Streptococcus sp, Staphylococcus aureus and 07 yeasts and fungi: Candida albicans, Candida spp, Penicillum spp, Aspegillus sp, Aspergillus flavus, Fusarium sp, and Cladosporium spp. The results of inhibition growth were compared with standard antimicrobial drugs with the goal of exploring their potential antimicrobial activity. In addition, the anti-inflammatory activity of the synthesized compounds was determined in-vitro by protein denaturation method. The obtained bioactivity results were further validated by in silico DFT (Density Functional Theory), ADME (Absorption-Distribution-Métabolisation-Excrétion), molecular docking studies and molecular dynamics simulations. Communicated by Ramaswamy H. Sarma

P339 A study of environmental risk factors in chronic rhinosinusitis

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM ObjectivesChronic rhinosinusitis (CRS) is a complex disease that incorporates many different conditions. Our aim is to determine the role of environmental risk factors in chronic rhinosinusitis.MethodsThis was a case-control study where forty cases of CRS in the age group of 18-62 years were recruited from the ENT-OPD of UCMS and GTB Hospital, Delhi, a tertiary care hospital, along with 40 age-appropriate healthy controls. Informed and written consent was obtained from all the subjects. Detailed history and examination along with routine blood and radiological investigations were done. Subjective assessment of disease was done using visual analog scales of SNOT 22 questionnaire, environmental questionnaire, and saccharine test. Objective assessment was done using endoscopic grading according to Lund and Kennedy scoring. CT scores were also obtained using Lund and Mackay staging system. Passive air sampling was done by the settle plate method and Anderson six-stage cascade impactor was used for active air sampling. Blood agar and Sabouraud dextrose agar plates were inserted on the six stages of the cascade impactor, which were later incubated at 35°C and 25°C respectively, for 18-24 hours. Bioaerosol concentrations were calculated and MicroScan WalkAway system was used to identify bacterial and fungal growth. The correlation between total bioaerosol concentration and the symptom score was done by spearman's correlation coefficient; a comparison of mean total bioaerosol concentration between the case and control groups was done by Mann-Whitney U-test.ResultsThe mean total bio-aerosol concentration from active air sampling in the case group was 458.52 CFU/m3 and 437.63 CFU/m3 in control, which was not significant. It was found to be negatively correlated, but not significant with various parameters in case groups like SNOT 22 score, environmental exposure score, saccharine test, smoking, and endoscopic score. A positive but insignificant correlation was found between total bioaerosol concentration and absolute eosinophilic count and CT score. The mean CFU/100 mm3 counted after passive air sampling between case and control group was 34.03 and 30.88 CFU/100mm3 which was not significant. Fungal growths were mostly Aspergillus flavus and A. Niger followed by Cladosporium spp.ConclusionThe fungal burden was high in all the households of the case as well as control groups, although, a significant correlation could not be established. The environmental conditions found in this study were found to be conducive to future risk of exacerbation of preexisting fungal infections for which, indoor air quality needs to be monitored.

S1.5b The burden of mycotic keratitis in West Africa

S1.5 Mycotic keratitis, September 21, 2022, 11:00 AM - 12:30 PM BackgroundFungal infection of the cornea, known as mycotic keratitis, can cause permanent corneal scarring and perforation resulting in the loss of the eye. This paper reviews the prevalence and epidemiology of mycotic keratitis in different countries in West Africa to estimate its burden.MethodsAn exhaustive search of the literature was made on Google, PubMed, MEDfacts, Cochrane Library, and Web of knowledge using different sets of keywords, viz. mycotic keratitis, ocular fungal infection, West Arica, risk factor, prevention, etc.ResultsA study in Nigeria over a period of 4 years (1974-1977) dealt with 42 confirmed cases of mycotic keratitis with Fusarium solani as the predominant etiological agent (14 cases) followed by Penicillium citrinum (8 cases) and Aspergillus fumigates (5 cases), Candida spp (3 cases). The remaining 12 cases were that of Fusarium moniliforme, Aspergillus spp, Penicillium sp, and Cladosporium spp. The predisposing factors identified were trauma from palm tree leaves, thorns, kernels, or other plant objects, mechanical tools, and frying oil. A 10-year review (2003-2012) of 152 cases of corneal ulcers at the University of Calabar Teaching Hospital, Calabar, Nigeria revealed only 2 (2.9%) cases due to Aspergillus sp, many patients in this study were farmers. Other studies from Nigeria only mentioned the prevalence of keratitis without any mention of fungal etiology. Of the two studies from Ghana, the one conducted in 1999 showed the predominant agents Fusarium spp. (52.3%) and Aspergillus spp. (15.3%), in the other one conducted in 1999-2001, these agents were represented by 42.2% and 17.4% respectively. In another prospective study of suppurative corneal ulcers in 290 cases in Ghana (June 1999-May 2001), the etiological agents identified in culturally proven 77 (85.5%) cases of mycotic keratitis were Fusarium spp-46, A. flavus-9, A. fumigates-7, A. niger-1, A. nidulans-1, and Aspergillus sp-1 A Siera Leonian study of cases of suspected infectious ulcerative keratitis from January 2005 to January 2006) detected 35.6% of mycotic keratitis and 13.7% of mixed fungal and bacterial etiology. A study on the burden of serious fungal infections in Togo mentioned an annual incidence of 951 cases of mycotic keratitis but no details of fungal etiology were mentioned.ConclusionInvestigators have estimated the annual global incidence of fungal keratitis at over 1 million cases. Reports of cases reported from some countries represent only a tip of the true burden of mycotic keratitis in West Africa. There is a need for comprehensive surveys (involving collaboration between ophthalmologists and microbiologists) of mycotic keratitis in representative communities in collaboration with primary health centers and hospitals in different countries. It should be possible to produce a combined antifungal antibacterial preparation for widespread and immediate prophylactic first-aid use after corneal trauma, especially in rural areas.

Cultivable microbiota of Proteus anguinus from underground habitats and animals accidently washed to the surface in Croatia

Proteus anguinus (olm) is an amphibian endemic to underground cave systems of the Dinaric karst of Central Europe, adapted to a life in complete darkness. Recent studies have demonstrated the global decline of amphibians due to poorly understood microbiological diseases, emphasizing just how little is known about the natural microflora of amphibians. Not much is known about the olm’s microbiota in their cave habitats, and nothing is known about the microbiology of animals washed to the surface. This observational study describes the microbiological analysis of the skin, cloaca and oral cavity of six olms collected from their cave habitats, and 16 collected after being washed out by flows from two springs, to learn more about the olm’s normal microbiota and possible changes after contact with the different environmental conditions on the surface. Standard microbiological procedures, MALDI-TOF and Real-Time PCR were used for microbiological species identification. All animals tested negative for Ranavirus, Batrachochytrium dendrobatidis, Batrachochytrium salamandrivorans and Chlamydia spp. The most abundant fungi isolated were Penicillium spp. and Cladosporium spp. The washed out animals showed a higher diversity of bacterial flora than those from cave habitats, with Acinetobacter johnsonii, Aeromonas hydrophila, Bacillus spp. and Janthinobacterium lividum as the most frequently identified isolates.

Metagenomic detection of eumycetoma causative agents from households of patients residing in two Sudanese endemic villages in White Nile State.

Eumycetoma is a chronic debilitating fungal disease endemic to tropical and subtropical regions, with Sudan featuring the highest eumycetoma incidence. Among the 50 species of fungi most commonly associated with eumycetoma Madurella mycetomatis (M. mycetomatis) is often referenced as the most common pathogen. However, there is an enormous knowledge gap related to this neglected disease and its pathogenesis, epidemiological features, and host-specific factors that could contribute to either the host susceptibility and resistance. In this study, we were able to utilize a metagenomic approach and samples collected from clinical black grains (BG) and familiar household environments aimed to assay both the habitat of eumycetoma-associated fungi and its possible connection with eumycetoma patients living in two different eumycetoma endemic villages within the White Nile State of Sudan. DNA sequencing targeting the fungal ITS2 domain was performed on soil, animal dung, housing walls and roofs, and Acacia-species thorn samples and compared with culture-dependent methods of fungal isolation. Additionally, we compared the soil samples obtained in the endemic zone with that from non-endemic zones, including Wagga village in Kassala State and Port Sudan suburb in Port Sudan State. Overall, a total of 392 Amplicon Sequence Variants (ASVs) were detected by ITS2 metagenomics Eumycetoma causative organisms accounted for 10% of total ASVs which included 11 genera: Exserohilum (2%), Aspergillus (1.7%), Curvularia (1%), Alternaria (0.9%), Madurella (0.5%), Fusarium (0.4%), Cladosporium (0.2%) Exophiala (0.15%), and, in a lesser extent, Microascus (0.05%) Bipolaris and Acremonium (0.01%) for each. Only five genera were identified by culture method, which included Fusarium (29%), Aspergillus (28%), Alternaria (2.5%), Bipolaris (1.6%), and Chaetomium (0.8%). M. mycetomatis was detected within all the studied patients' houses, accounting for 0.7% of total sequences. It was the first common eumycetoma-associated agent detected in soil samples and the third common in the dung and wall samples. In contrast, it was not detected in the roof or thorn samples nor in the soils from non-endemic regions. Exserohilum rostratum, Aspergillus spp and Cladosporium spp were detected in all samples. M. mycetomatis and other eumycetoma-associated fungal identified in the patients' black grains (BG) samples by metagenomics were identified in the environmental samples. Only Acremonium alternatum and Falciformispora senegalensis, responsible for eumycetoma in two patients were not detected, suggesting the infections in these patients happened outside these endemic areas. The soil, animal dung, and houses built from the same soil and dung are the main risk factors for M. mycetomatis infection in these endemic villages. Furthermore, the poor hygienic and environmental conditions, walking barefooted, and the presence of animals within the houses increase the risk of M. mycetomatis and other fungi causing eumycetoma.

Antioxidant and Antibacterial Activity of Bioactive Compound Produced By Enophytic Fungi

The present study aims evaluated the antibacterial and antioxidant activity of bioactive compounds synthesized by endophytic fungi obtained from Anacardium occidentale (AO), Terminalia cattappa (TC) and Sygygium jambos (SJ). The compounds produced by the isolated endophytic fungi from the medicinal plants used in this study were extracted with ethylacetate and subjected to antioxidant and antibactrerial activities using standard procedures. Screened for bioactivity using the disc diffusion method. Five fungal species were isolated from the plants and were identified to be Aspergillus terreus, Cladosporium spp, Mucor racemosus, Aspergillus oryzae, F. oxysporum. Alkaloids, saponins and steroids were detected in all the extracts while tannin and phenols were not detected. However, flavonoids was detected in SJ1 and TC2 only. All the extracts obtained from this study showed good antioxidant activity with SJ1 having the highest DPPH radical scavenging compared to the other extracts. All the extracts inhibited the growth of both E. coli and S. aureus with a range of 10-35 mm in diameter. The MBC: MIC of AO, SJ1 and SJ2 was 2.0 for both S. aureus and E. coli while that of TC2 was 10. In conclusion, the compounds produced by the isolated fungi possess both antioxidant and antibacterial activity which can be used for development of new drugs.

Defining Mycotoxins Associated with Wheat Grains in Mosul Silo by ELISA

The study aimed to identify fungi-contaminated with some varieties of wheat grains locals, as well as isolate fungi-contaminated with grains imported wheat in Silo of Mosul city, and estimate the level of mycotoxins ochratoxin, zearalenone, and aflatoxin in them by ELISA method. Results showed that the local wheat variety, Ebaa 99, showed the highest percentage of appearance in Aspergillus flavus, was 63.15%, while the lowest appearance of fungi was Mucor spp. With a percentage of 2.47%, the variety Rasheed showed the highest percentage of A. flavus, which was 49.12%, while the lowest appearance was of Rhizopus spp. With a percentage of 2.29%, As well as in the variety Al-Baraka A. flavus showed the highest percentage, which was 5.453%, and the lowest percentage was Cladosporium spp by 9.43%. The contaminated fungi differed from the imported varieties and the percentage of their appearance. In the Canadian variety, the highest appearance of A. flavus was 63.72%, and A. dematatus was the least appearance with 5.42%. The fungus A. flavus also appeared in the American variety, with the highest appearance of 43.17%. The least appearance was Fusarium oxysporum at 5.34%. In the Australian variety, the highest appearance was found in A. flavus, at 54.03%, and the fungus Fusarium spp. was the least appeared fungi, at 1.96%. Estimation of mycotoxins indicates that the aflatoxin amount in the variety Ebaa 99 samples was 34.55 ppm, while its amount of ochratoxin and zearalenone was 21.75 ppm and 0.23 ppm, respectively. Canadian variety grains had the lowest aflatoxin content, with the highest ochratoxin. Australian variety grains had the lowest content of aflatoxin, ochratoxin, and zearalenone amount 12.29 ppm, 0.82 ppm, and 11.51 ppm, respectively.