Penicillium Species Research Articles

A comparison of microbiota isolation methods reveals habitat preferences for fermentative yeasts and plant pathogenic fungi in the grape berry

The methodologies for profiling the grape berry microbiota have exponentially evolved in the past 25 years. Recently, concerns arose regarding the homogeneity in the protocols of grape harvesting, sequencing and bioinformatic analyses, but the bias introduced by the microbiota isolation method is still unexplored. This study followed a simple approach of comparing two most used methods of microbiota collection from grape berries (washing vs crushing), hypothesizing a significant impact in the outcome of the microbiota profiles analyzed by NGS metabarcoding. Experiments conducted in fruits of three cultivars of the Douro wine region showed that only 52 % of OTUs were common to both surface and juice microbiota, suggesting specific microbial niches. Thirteen fungal genera were abundantly detected in the fruit surface, including Alternaria, Aureobasidium, Cladosporium, Didymella and Bipolaris. Fermentative yeasts including Meyerozyma and Saccharomyces cerevisiae were exclusively detected in the juice, together with several Penicillium species. Distinct habitat preferences of species within the genera Alternaria, Sporobolomyces and Rhodotorula were also revealed. The study showed that the microbiota isolation method is crucial in the detection of certain plant pathogenic/saprophytic fungi and yeasts with biotechnological and oenological interest, adding novelty to the globally accepted assumption that S. cerevisiae in musts originates primarily from the cellar.

First identification of potato tuber rot caused by Penicillium solitum, its silver nanoparticles synthesis, characterization and use against harmful pathogens.

Potato is one of the highly consumed vegetable crop grown in different regions across Pakistan that is affected by fungal diseases. The current research was conducted to identify fungal pathogen causing mold-like disease of potato in Khyber Pakhtunkhwa (KP), Pakistan. For molecular identification and characterization of the fungal disease; potato tuber samples were collected followed by culturing on potato dextrose agar (PDA). Based on morphological features, the pathogen was identified as a Penicillium species. This result was obtained in 45 different isolates from potato tubers. Molecular identification was done using β-tubulin primers and ITS5 sequencing of 13 different isolates that releveled 98% homology with BLAST (GenBank accession no. KX958076) as Penicillium solitum (GenBank accession nos. ON307317; ON307475 and ON310801). Phylogenetic tree was constructed that showed Penicillium solitum prevalence along with Penicillium polonicum and Penicillium citrinum on potato tubers. Based on this, Penicillium solitum based silver nanoparticles (Ag NPs) were synthesized and characterized using UV-visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), energy dispersive X-ray (EDX) and field emission scanning electron microscopy (FE SEM). UV-analysis showed a characteristic peak at 410 nm confirming synthesis of Penicillium solitum based Ag NPs. This was further confirmed by XRD followed by EDX and SEM that showed face cubic crystal structure with Ag as major constituent of 18 nm formed spherical Ag NPs. FTIR showed band stretching of O-H, N-O and C-H of biological origin. Similarly, Penicillium solitum based Ag NPs presented strong anti-bacterial and anti-fungal activity at 0.5 level of significance LSD. According to our knowledge, this is the first report of Penicillium solitum identification in Pakistan, its Ag NPs synthesis and characterization to be used against pathogens of agricultural significance.

Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum

Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain of P. oxalicum was associated with enhanced expression of a cyclophilin gene, PoxCYP18. Cloning and characterization of PoxCYP18 revealed that its cDNA consists of 522 bp encoding a protein of 173 amino acid residues, with predicted molecular mass and pI values of 18.91 kDa and 8.87, respectively. The recombinant PoxCYP18 can catalyze cis–trans isomerization of peptidyl-prolyl bond with a catalytic efficiency of 1.46 × 107 M−1 s−1 and is inhibited specifically only by cyclosporin A, with an inhibition constant of 5.04 ± 1.13 nM. PoxCYP18 consists of two cysteine residues at positions − 45 and − 170, and loses its activity under oxidizing conditions. Substitution of these residues alone or together by site-directed mutagenesis revealed that the PPIase activity of PoxCYP18 is regulated through a redox mechanism involving the formation of disulfide linkages. Heterologous expression of PoxCYP18 conferred enhanced tolerance to salt stress in transgenic E. coli cells, implying that this protein imparts protection to cellular processes against salt-induced damage.

Endophytic Fungal Biodiversity and Antimicrobial Bioprospecting of Penicillium citrinum KIB4 from Khaya ivorensis Bark

Background: The emergence of pathogens that are resistant to antibiotics has raised serious concerns about public health, forcing the hunt for alternative antibacterial substances. Endophytes, particularly those from unscreened plants, may offer new sources of antibacterial substances. Hence, this research aimed to isolate antibiotic-producing fungi from Khaya ivorensis bark, a medicinal plant widely used in Nigeria. Methods: The fungal endophytes were isolated following conventional methods using potato dextrose agar (PDA). They were characterized by morpho-microscopical and sequencing of the internal transcribed spacer (ITS) of the fungal rDNA. The phylogenetic tree was drawn following the Neighbour-joining method and the tree was annotated using the Interactive Tree of Life (iTOL) version 6.0. The agar plug assay method was used to select putative antibiotic-producing isolate for further research. The antimicrobial activity of the fungal fermentation product was assessed by agar well diffusion while the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were estimated using the solid agar dilution technique. Results:Atotal of 12 endophytic fungi were isolated. They were noted to be mostly Aspergillus sp. (11 isolates) while only one isolate represented Penicillium species. Only isolate KIB4 showed good inhibitory activity against several bacterial pathogens tested in the preliminary study within 10 mm to 12 mm inhibition zone diameter (IZD) range. Molecular identification proved that strain KIB4 fell into the genus Penicillium and shared the highest ITS gene sequence similarity with Penicillium citrinum D112 (98.83%), and Penicillium sp. TWJL1743 (98.65%). The fermentation extract of Penicillium citrinium KIB4 showed specific inhibition against Escherichia coli ML475, Staphylococcus aureus LN001, Klebsiella pneumoniae ML602, Bacillus subtilis ML225, and Proteus sp. UR979 with an IZD range of 11mm to 23mm. Conclusion: We reported for the first time, the antimicrobial efficacy of fermentation extract from endophytic fungi isolated from Khaya ivorensis against several bacterial pathogens. Further fractionation of the crude extract is needed to isolate specific antimicrobial compounds.

Molecular Characterization of Fungal Isolates of Locally Processed Rice from Five Agro-Ecological Zones of Nigeria

The purpose of this study was to characterize fungal isolates contaminating locally processed rice in Nigeria and to assess the genetic similarity in the fungi isolates. Fungal species were characterized by amplification of a segment of the β-tubulin gene using Bt2aF (5’-GGTAACCAAATCGGTGCTGCTTTC-3’) and Bt2bR (5’ACCCTCAGTGT GT GACCCTTGGC-3’) primers and phylogenetic relationship established using the Maximum Likelihood method. Amplification of between 450 and 550bp of the β-tubulin region of the genomic DNA of the fungal isolates was obtained from thirty-eight samples (89%). Members of Aspergillus section flavi and section nigri groups were differentiated as A. flavus, A. parasiticus, A. tubigensis, A. sydowii and A. niger respectively. Molecular method identified Paecilomycesformosus with 99% similarity index while all the morphologically identified Penicillium species were identified to genus level as P. formosus, P. steckii and P. citrinum with a percentage similarity ranging from 95%-100%. Collectotrichum fruticola and Talaromyces flavus from SGS and NGS zone respectively were documented for the first time in the locally processed rice with 100% and 89% similarity index respectively. All the Aspergillus species were closely related to one another and narrowly diverged from the Penicillium species. Aspergilla and Penicillia were the predominant genera identified and the isolates were genetically similar irrespective of their agroecological zones.

Safety Concerns on Microbes Associated with Fresh and Smoked Fish Sold in Igbokoda Fish Market, Nigeria

Fishes are important source of food for human globally because man gets a lot of minerals, vitamins, lipids and proteins from fishes and theirs products. Microbiological quality of fish is important to public health as indicative parameters to ensure safe consumption of fish by man. This study was carried out in order to evaluate microbes associated with fresh and smoked fishes on sales at Igbokoda fish market and their antimicrobial susceptibility profiles. Ten different fresh and smoked fish samples were collected from different fish-sellers at different selling points in Igbokoda fish market, aseptically using sterile containers. Samples from the skin, intestine and gill were obtained and cultured into three different media (MacConkey Agar, Nutrient Agar, Salmonella-Shigella Agar) for bacteria isolation and characterization. Other samples from the skin, intestine and gill were also cultured into Potato Dextrose Agar for fungal isolation and characterization. After 48 hours, on examination the total bacterial counts obtained from the skin, intestine and gill ranged between 2.4 to 8.7 x 106 cfu/ml while the total fungal counts ranged between 4.6 to 9.2 x 104 cfu/ml. Highest microbial load was obtained from the skin of the fish samples (7.2 x 106 cfu/ml), while the gill had the lowest microbial load (4.6 x 104 cfu/ml). The bacteria species isolated from the fish samples and their percentage of occurrence were; Aeromonas hydrophilia (11.2%), Bacillus species (6.8%), Citrobacter freundii (3.3%), Escherichia coli (23.2%), Enterobacter cloacae (6.1%), Enterococcus faecalis (6.5%), Listeria monocytogenes (4.5%), Pseudomonas aeroginosa (8.2%), Salmonella species (10.2%), Staphylococcus aureus (16.5%) and Streptococcus species (3.5%). The fungi isolated and their percentage of occurrence were; Penicillium species (16.2%), Aspergillus flavus (8.4%), Aspergillus niger (25.5%), Candida species (16.4%), Fusarium species (6.5%), Mucor species (8.6%), Rhodotorula species (7.2%) and Rhizopus stolonifer (11.2%). Antibiotics susceptibility testing showed that all bacteria isolates were susceptible to Chloramphenicol, Ciprofloxacin, Augmentin, and Amoxillin. While Salmonella species, Staphylococcus aureus and Streptococcus species showed strong resistance to Septrin, Gentamycin and Streptomycin but intermediate susceptible to Erythromycin, Ampiclox, and Tetracycline. The antifungal susceptibility testing of the fungal isolated from fish samples showed that all the isolates were susceptible to all the antifungal agents used except Candida species that showed little resistance to clotrimazole. The study showed that microbial loads in the fish samples were low and, also there was no presence of multidrug resistant bacteria in sampled fish; therefore the fishes were safe for consumption. Although, it is recommended that good processing method should be adopted by fish handlers to prevent health risks to the fish consumers.

Evaluation of the Antibiotic Susceptibility Patterns of Isolates from Feed and Water of Selected Poultry Farms in Awka Anambra State, Nigeria

The susceptibility of microorganisms isolated from poultry feeds and poultry water samples to selected antibiotics was assessed. Standard methods were used to analyze selected poultry feeds and poultry water samples. The antibiotic susceptibility patterns of the bacterial isolates were determined against the following antibacterial agents: erythromycin (10 µg), ciprofloxacin (10 µg), ampiclox (20 µg), rifampicin (20 µg), amoxil (20 µg), septrin (30 µg), ampicillin (30 µg), ceporex (10 µg), levofloxacin (20 µg), gentamicin (10 µg), streptomycin (30 µg), norfloxacin (10 µg), chloramphenicol (30 µg), ofloxacin (10 µg), nalidixic acid (30 µg), reflecine (10 µg), and augmentin (30 µg).The highest viable counts of bacteria isolated from poultry feed and water samples were 2.7x106 cfu/g and 1.69x103 cfu/ml, respectively. The highest fungal counts in the poultry feed and water samples were 1.60x105 cfu/g and 2x105 cfu/ml, respectively. Bacterial isolates from poultry feed and water samples included Escherichia coli, Klebsiella pneumoniae, Salmonella species, and Staphylococcus aureus. Fungal isolates included Aspergillus species, Penicillium species, Mucor species, and Candida species.Staphylococcus aureus exhibited the highest susceptibility to most of the antibiotics, while Klebsiella pneumoniae showed the highest resistance, as it was resistant to five out of the ten antibiotics tested in this study. The research has demonstrated that poultry feed and poultry water showed varying levels of contamination, which may pose serious health risks to poultry. Amoxil, levofloxacin, ciprofloxacin, reflecine, and ofloxacin are recommended for use as antibiotics to treat diseases that may be caused by some of these pathogens.

Regional distribution and diversity of Aspergillus and Penicillium species on Croatian traditional meat products

Various factors, such as weather and production practices (e.g., environmental hygiene, process duration, raw material quality, ripening temperature, and relative humidity), in combination with the intrinsic product properties (e.g., pH, aw, salt content), significantly affect the growth of surface moulds. The aim of this study was to isolate and identify surface moulds retrieved from traditional meat products (TMPs) and correlate these data to the production region and production technology. The surface of 250 TMPs (dry-fermented sausages, n = 108; dry-cured meat products, n = 142) from five Croatian regions were sampled during a two-year period. Dry-fermented sausages had a significantly higher pH and a lower salt concentration when compared to dry-cured meat products. In total, 528 isolates were obtained, comprising 20 Penicillium and 17 Aspergillus species. The species most frequently isolated from the dry-fermented sausages were P. commune (32.4 %), A. proliferans (33 %), and P. solitum (14.8 %), while A. proliferans (52.1 %), P. commune (28.9 %) and P. citrinum (19.7 %) predominated in dry-cured meat products. Aspergillus predominated on the TMPs from southern Croatia, while Penicillium was prevalent on products from the other four regions, possibly due to differences in weather conditions. Seven potentially mycotoxigenic species (A. creber, A. flavus, A. niger, A. westerdijkiae, P. citrinum, P. commune, and P. nordicum) were isolated and identified. Regular monitoring of mould species and their toxigenic metabolites present on traditional meat products is of the utmost importance from the public health perspective, while the results of such a monitoring can prove beneficial for the tailoring of the production technology development.

Mycobiota in Slovak wine grapes: A case study from the small Carpathians wine region.

The microbiological characteristics of the grapes are made up of a wide variety of microorganisms, including filamentous fungi. Their presence in grapes is traditionally associated with deterioration in quality. The health of the grapes is very important for obtaining quality wine. The objective of this study was to investigate the diversity of mycobiota on the surface and inside of different grapevine varieties at harvest time in the temperate climate of Slovakia and to identify potentially pathogenic isolates of Aspergillus and Penicillium producing selected mycotoxins. During the 2021 grape harvest, grapes were collected from the Small Carpathians wine region. Eleven grape samples were analyzed by the plating method and plating method with surface disinfection. Emphasis was placed on Aspergillus and Penicillium species because of their importance in mycotoxin production. Of the 605 fungal strains detected, 11 genera were identified in the exogenous mycobiota. The most common and abundant genera were Alternaria and Botrytis. In the genus Aspergillus, A. section Nigri is the most abundant, while in the genus Penicillium, P. raistrickii reached the highest frequency and abundance. Of the 379 strains detected and identified from the endogenous mycobiota, the most common genera were again Alternaria and Botrytis and the most abundant genus was Botrytis. Penicillium species were detected in 17% of all fungi found, with P. raistrickii dominating. The A. section Nigri reached only 4% of the relative density of all isolates. Potentially toxigenic Aspergillus and Penicillium species were tested for toxinogenity by thin layer chromatography. The most important mycotoxin-producing species found were A. section Nigri but without ochratoxin A production.

Assessment and Biodegradation of Polycyclic Aromatic Hydrocarbons in Soil and Water Around Petroleum Products Depot Suleja, Nigeria.

Petroleum contamination constitutes a frequent incidence in various petroleum depots in Nigeria. In this study, the polycyclic aromatic hydrocarbons (PAHs) present in soil and water in communities around Petroleum Products Marketing Company (PPMC) Suleja, Nigeria, were evaluated and degraded using indigenous microorganisms. The samples sites were divided into 7 plots from where samples of water and soil were obtained: one within the PPMC depot, five from communities surrounding the depot, and the control 93,000km from the depot. The microbial counts were determined using spread plate inoculation technique on minimal salt media. The microbial isolates were characterized and identified based on their cultural, biochemical, and molecular characteristics. The potential of the microbial isolates to utilize 0.05 mL of diesel, kerosene, engine oil, and crude oil was determined in a Bushnell Haas Broth, and the biodegradation was determined by total viable cell counts and spectrophotometry. The ability of the isolates to mineralize PAHs was also evaluated in a minimum salt media. The bacterial isolates were species of Streptococcus, Pseudomonas, Staphylococcus, Proteus, Escherichia, and Bacillus, while species of Penicillium, Aspergillus, Mucor, and Rhizopus were isolated among the fungi. Aspergillus niger strain ATCC 1015 and Bacillus thuringiensis strain M43 showed high capacity to utilize the 16 priority PAHs. The pahE1 gene was used by Bacillus thuringiensis, Pseudomonas aeruginosa and A. niger, while Penicillium notatum used pahE2 gene for the degradation of the PAH. The current study identified microbial isolates that can utilize priority PAHs, making them beneficial for oil spill bioremediation in tropical environments.

Effect of phyllosphere and spike mycoflora on castor gray mold incited by Amphobotrys ricini

Gray mold of castor incited by Amphobotrys ricini is an economically important disease causing heavy lossesduring capsule formation stage under high humid conditions. The phyllosphere and spike mycoflora were isolated from castor and tested for their bioefficacy against gray mold under laboratory and greenhouse conditions. The mycoflora isolated were identified based on morphological characters as species of Acremonium, Aspergillus, Choanephora, Cladosporium, Fusarium, Macrophomina and Penicillium. Among the mycoflora tested in vitro, highest inhibition of the test pathogen was recorded with Acremonium species. Greenhouse evaluation of phyllosphere and spike mycoflora by detached spike method has shown that inoculation of detached spikes with Penicillium spp. 24 hours prior to pathogen inoculation has significantly reduced gray mold severity compared to control.

Co-occurrence of mycotoxins and other fungal metabolites in total mixed rations of cows from dairy farms in Punjab, Pakistan

After India and the USA, Pakistan is the third country leading in global dairy production, a sector of very high socioeconomic relevance in Asia. Mycotoxins can affect animal health, reproduction and productivity. This study analysed a broad range of co-occurring mycotoxins and fungal secondary metabolites derived from Alternaria, Aspergillus, Fusarium, Penicillium and other fungal species. To complete this, a validated multi-metabolite liquid chromatography/electrospray ionization-tandem mass spectrometric (LC/ESI–MS/MS) method was employed, detecting 96 of > 500 tested secondary fungal metabolites. This first preliminary study demonstrated that total mixed rations (TMRs) (n = 30) from big commercial dairy cattle farms (> 200 lactating cows) in Punjab, Pakistan, presented ubiquitous contamination with mixtures of mycotoxins. The mean of mycotoxins per sample was 14, ranging from 11 to 20 mycotoxins among all TMR samples. Metabolites derived from other fungi and Fusarium spp. showed the highest levels, frequency and diversity among the detected fungal compounds. Among the most prevalent mycotoxins were Fusarium toxins like fumonisins B1 (FB1) (93%), B2 (FB2) (100%) and B3 (FB3) (77%) and others. Aflatoxin B1 (AFB1) was evidenced in 40% of the samples, and 7% exceeded the EU maximum limit for feeding dairy cattle (5 µg/kg at 88% dry matter). No other mycotoxin exceeds the EU guidance values (GVs). Additionally, we found that dietary ingredients like corn grain, soybean meal and canola meal were related to increased contamination of some mycotoxins (like FB1, FB2 and FB3) in TMR from the province of Punjab, Pakistan. Among typical forage sources, the content of maize silage was ubiquitous. Individually, the detected mycotoxins represented relatively low levels. However, under a realistic scenario, long-term exposure to multiple mycotoxins and other fungal secondary metabolites can exert unpredictable effects on animal health, reproduction and productivity. Except for ergot alkaloids (73%), all the groups of metabolites (i.e. derived from Alternaria spp., Aspergillus spp., Fusarium spp., Penicillium spp. and other fungi) occurred in 100% of the TMR samples. At individual levels, no other mycotoxins than AFB1 represented a considerable risk; however, the high levels of co-occurrence with several mycotoxins/metabolites suggest that long-term exposure should be considered because of their potential toxicological interactions (additive or synergistic effects).

Domestication in dry-cured meat Penicillium fungi: Convergent specific phenotypes and horizontal gene transfers without strong genetic subdivision.

Some fungi have been domesticated for food production, with genetic differentiation between populations from food and wild environments, and food populations often acquiring beneficial traits through horizontal gene transfers (HGTs). Studying their adaptation to human-made substrates is of fundamental and applied importance for understanding adaptation processes and for further strain improvement. We studied here the population structures and phenotypes of two distantly related Penicillium species used for dry-cured meat production, P. nalgiovense, the most common species in the dry-cured meat food industry, and P. salamii, used locally by farms. Both species displayed low genetic diversity, lacking differentiation between strains isolated from dry-cured meat and those from other environments. Nevertheless, the strains collected from dry-cured meat within each species displayed slower proteolysis and lipolysis than their wild conspecifics, and those of P. nalgiovense were whiter. Phenotypically, the non-dry-cured meat strains were more similar to their sister species than to their conspecific dry-cured meat strains, indicating an evolution of specific phenotypes in dry-cured meat strains. A comparison of available Penicillium genomes from various environments revealed HGTs, particularly between P. nalgiovense and P. salamii (representing almost 1.5 Mb of cumulative length). HGTs additionally involved P. biforme, also found in dry-cured meat products. We further detected positive selection based on amino acid changes. Our findings suggest that selection by humans has shaped the P. salamii and P. nalgiovense populations used for dry-cured meat production, which constitutes domestication. Several genetic and phenotypic changes were similar in P. salamii, P. nalgiovense and P. biforme, indicating convergent adaptation to the same human-made environment. Our findings have implications for fundamental knowledge on adaptation and for the food industry: the discovery of different phenotypes and of two mating types paves the way for strain improvement by conventional breeding, to elucidate the genomic bases of beneficial phenotypes and to generate diversity.

Isolation and Identification of Mold Contaminants in Peanut Paste from the Cengkareng Market of West Jakarta

The filamentous molds associated with peanut paste produce biochemicals known as mycotoxins. The major mycotoxins are aflatoxins produced by molds from the genera Aspergillus and Penicillium. Mold contaminants can be occurred due to poor hygiene storage. The purpose of this study was to determine the presence of molds in peanut paste sold at the Cengkareng market in order to assist people in being attentive to the storage conditions of peanut paste. Samples were collected randomly with a total of 15 packages of peanut paste from Cengkareng market, West Jakarta, Indonesia. Potato Dextrose Agar was used for the isolation and identification of molds. The molds were observed based on the morphological features through macroscopic and microscopic examination. The frequencies of 15 samples contaminated with molds were computed on a percentage. From the observed morphological features and colony colour of molds, those were A. niger, A. flavus, Penicillium sp, and Mucor sp. This research also revealed that of the 15 samples identified, A. niger and A. flavus, 40% respectively. Other species Penicillium sp. 27% and Mucor sp. 13% were the least common. Therefore, this study confirmed that Aspergillus is the highest risk of contaminating peanut paste compared to other genera, which can cause a health problem for humans.

Microbiology and Physicochemical Characteristics of Surface Water from a Local Refinery Site in Port Harcourt, Nigeria

Water plays a vital role in maintaining human health but due to an increase in industrialization, urbanization and other anthropogenic activities, surface water pollution is on an increase. This research was carried out to investigate the microbiological and physicochemical status of a crude oil polluted surface water in Iwofe River, Rumuolemini, Port Harcourt. Samples were collected from two points (Upstream and Downstream) bi-monthly for a period of three months. The THB counts for the upstream and downstream samples were 1.7 × 108cfu/ml and 1.4 × 108 cfu/ml, respectively while the THF counts for the samples were 3.0 × 106cfu/ml and 1.4 × 106 cfu/ml, respectively. Hydrocarbon utilizing bacterial (HUB) counts were higher in the upstream (1.5 × 106cfu/ml) than downstream (4.3 ×105cfu/ml) location of the Iwofe River. The Hydrocarbon utilizing fungal (HUF) count ranged from 1.0 × 105cfu/ml to 1.4 × 105 cfu/ml in the upstream and downstream water samples, respectively. The results of all the microbiological parameters analyzed indicated that the total heterotrophic bacterial (THB) and Total heterotrophic fungal (THF) counts were higher in the upstream than in the downstream location. The Most Probable Number (MPN) method used for the determination of coliform counts revealed the presence of coliforms both upstream and downstream, with an MPN index of 1,600/100ml of each sample. Some of the bacterial species isolated were Staphylococcus aureus, Escherichia coli and Vibrio cholerae while the fungal isolates included Aspergillus fumigatus, Penicillium species and Alternaria species. The high level of oil contamination poses a great health threat and there is a need for remediation of the aquatic ecosystem.

Endophytic fungal community of Rosa damascena Mill. as a promising source of indigenous biostimulants: Elucidating its spatial distribution, chemical diversity, and ecological functions

The role of endophytes in maintaining healthy plant ecosystems and holding promise for agriculture and food security is deeply appreciated. In the current study, we determine the community structure, spatial distribution, chemical diversity, and ecological functions of fungal endophytes of Rosa damascena growing in the North-Western Himalayas. Culture-dependent methods revealed that R. damascena supported a rich endophyte diversity comprising 32 genera and 68 OTUs. The diversity was governed by climate, altitude, and tissue type. Species of Aspergillus, Cladosporium, Penicillium, and Diaporthe were the core endophytes of the host plant consisting of 48.8% of the endophytes collectively. The predominant pathogen of the host was Alternaria spp., especially A. alternata. GC-MS analyses affirmed the production of diverse arrays of volatile organic compounds (VOC) by individual endophytes. Among the primary rose oil components, Diaporthe melonis RDE257, and Periconia verrucosa RDE85 produced phenyl ethyl alcohol (PEA) and benzyl alcohol (BA). The endophytes displayed varied levels of plant growth-promoting, colonization, and anti-pathogenic traits. Between the selected endophytes, P. verrucosa and D. melonis significantly potentiated plant growth and the flavonoids and chlorophyll content in the host. The potential of these two endophytes and their metabolites PEA and BA was confirmed on Nicotiana tabacum. The treatments of the metabolites and individual endophytes enhanced the growth parameters in the model plant significantly. The results imply that P. verrucosa and D. melonis are potential plant growth enhancers and their activity may be partially due to the production of PEA and BA. Thus, R. damascena harbors diverse endophytes with potential applications in disease suppression and host growth promotion. Further investigations at the molecular level are warranted to develop green endophytic agents for sustainable cultivation of R. damascena and biocontrol of leaf spot disease.

The air and dust invisible mycobiome of urban domestic environments

Air and dust harbor a dynamic fungal biome that interacts with residential environment inhabitants usually with negative implications for human health. Fungal air and dust synthesis were investigated in houses across the Athens Metropolitan area. Active and passive culture dependent methods were employed to sample airborne and dustborne fungi for two sampling periods, one in winter and the other in summer. A core mycobiome was revealed both in air and dust constituted of the dominant Penicillium, Cladosporium, Aspergillus, Alternaria and yeasts and accompanied by several common and rare components. Penicillium and Aspergillus diversity included 22 cosmopolitan species, except the rarely found Penicillium citreonigrum, P. corylophilum, P. pagulum and Talaromyces albobiverticillius which are reported for the first time from Greece. Fungal concentrations were significantly higher during summer for both air and dust. Excessive levels of inhalable aerosol constituted mainly by certain Penicillium species were associated with indoor emission sources as these species are household molds related to food commodities rot. The ambient air fungal profile is a determinant factor of indoor fungal aerosol which subsequently shapes dustborne mycobiota. Indoor fungi can be useful bioindicators for indoor environment quality and at the same time provide insight to indoor fungal ecology.

Health Implication of Mycotoxigenic Fungi and Mycotoxins Analysed from Sun Dried Yam and Potato Chips Sold in Minna Niger State, Nigeria

Mycotoxin is toxic secondary metabolite synthesized by filamentous fungi during their stationary phase of growth in agricultural crops, food and feed products. Niger State experienced high temperature and humidity for about three-quarters of the year, and these are some of the conditions that encourage filamentous fungi to produce mycotoxin in food products. Enzyme Linked Immunosorbent Assay (ELISA) was used to dectect mycotoxins. Mycological profile and mycotoxins of sun dried yam and potato chips was determined. Total of fifty (50) samples were collected randomly from five markets in Minna and its environs. 179 isolates were recorded. Aspergillus species 86(53.1 %) Mucor species 27(15.1 %), yeast species 25(13.9 %) Penicillium species 14(7.8 %), Fusarium species 14(7.8 %) Rhizopus species 3(1.7 %) and Microsporum species 1(0.6 %) were the common fungi. The highest mean fungal counts (12.8 x 103 cfu/g) were observed in sweet potato chips bought in Gwada market while the lowest (3.5 x 103 cfu/g), was observed in Yam chips from Paiko market. The mean moisture content of 7.41±2.28 % was recorded in potato chips and yam chips had the lowest moisture content of 4.72± 1.66 %. Total aflatoxins, fumonisin and zearalenone contents were detected in 48 (96 %) of the samples. In yam and potato chips, total aflatoxin contamination was 68 and 84 % with mean concentrations of 12.6±7.36 μg/kg and 10.3±13.81 μg/kg respectively. The total aflatoxin concentration range of yam and potato chips were 0-61.84 μg/kg and 0-54.49 μg/kg with 28 and 48 % of the samples below 4μg/kg E.U limit respectively. Fumonisin contamination was 80 and 60% with mean concentrations of 928.123±559.50 μg/kg and 722.64 ± 137.85 μg/kg with a range 0.0-4018.24 μg/kg and 0 – 2055.98 μg/kg and had 52 and 72 % below 1000 μg/kg acceptable limit for yam and potato chips respectively. Zearalenone contamination in yam and potato account for 56 and 68% having mean concentration of 16.33± 9.71 μg/kg and 24.12± 21.03 μg/kg with a range 0 – 45.53 μg/kg and 0 – 83.74 μg/kg respectively. All the samples contaminated with Zearalenone were below the E.U regulatory limit of 100 μg/kg for food stuff intended for direct consumption. Dietary exposure and risk characterisation (EDI and %TDI) were further estimated for total aflatoxins, fumonisin and zearalenone. It was concluded that the level of contamination of these chips with mycotoxins poses a serious threat to public health and there is the need for proactive approach to drastically reduce fungal contamination.

Plant growth-promoting traits and activation of defense enzymes in traditional rice variety by fungal endophytes isolated from seeds of indigenous rice cultivars of Northeast India

The present investigation was carried out to expound on the role of seed borne endophytic fungi isolated from native rice cultivars of North East India. Altogether 226 fungal isolates were obtained from the surface-sterilized seeds of the 10 selected rice cultivars. The major isolates were species of Curvularia, Aspergillus, Fusarium, Gliocladium, Mucor, Penicillium, Bipolaris, Basidiobolus, Periconia, and non-sporulating mycelia sterilia. Principal Component Analysis (PCA) revealed dominance of Fusarium and Curvularia with high degree of colonization in Ampack and Chakhao rice cultivars. Altogether 84.61% of the isolates, showed siderophores activity on CAS agar medium and 76.92% of the isolates showed phosphorus solubilization activity. The result indicated that 84.61% of the isolates showed positive for ammonia production and interestingly grew in nitrogen-deficient Jensen's agar medium. The fungal isolates BD9, KON7, AM34, AM10, and Mycelia sterilia exhibited the highest antagonistic activity against rice pathogen Magnaporthe grisea. Enhanced production of peroxidase, polyphenol oxidase, chitinase, and superoxide dismutase enzyme was observed when a traditional rice plant was inoculated with a consortium made of fungal isolates AM34 and AM10 (Treatment 3). This combination exhibited a moderate increase in catalase production. Consortia made of other fungal isolates were not efficient in producing any defense enzymes. The isolates AM10, AM28, AM16, AM34, and AN 20 were identified as Fusarium annulatum, Daldinia eschscholtzii, Montagnula scabiosae, Fusarium redolens and Aspergillus flavus respectively based on microscopic and molecular characterization

Multiple Clonostachys rosea UDP-Glycosyltransferases Contribute to the Production of 15-Acetyl-Deoxynivalenol-3-O-Glycoside When Confronted with Fusarium graminearum.

Mycotoxins, derived from toxigenic fungi such as Fusarium, Aspergillus, and Penicillium species have impacted the human food chain for thousands of years. Deoxynivalenol (DON), is a tetracyclic sesquiterpenoid type B trichothecene mycotoxin predominantly produced by F. culmorum and F. graminearum during the infection of corn, wheat, oats, barley, and rice. Glycosylation of DON is a protective detoxification mechanism employed by plants. More recently, DON glycosylating activity has also been detected in fungal microparasitic (biocontrol) fungal organisms. Here we follow up on the reported conversion of 15-acetyl-DON (15-ADON) into 15-ADON-3-O-glycoside (15-ADON-3G) in Clonostachys rosea. Based on the hypothesis that the reaction is likely being carried out by a uridine diphosphate glycosyl transferase (UDP-GTase), we applied a protein structural comparison strategy, leveraging the availability of the crystal structure of rice Os70 to identify a subset of potential C. rosea UDP-GTases that might have activity against 15-ADON. Using CRISPR/Cas9 technology, we knocked out several of the selected UDP-GTases in the C. rosea strain ACM941. Evaluation of the impact of knockouts on the production of 15-ADON-3G in confrontation assays with F. graminearum revealed multiple UDP-GTase enzymes, each contributing partial activities. The relationship between these positive hits and other UDP-GTases in fungal and plant species is discussed.