Fungal Samples Research Articles

Synthesis, Characterization, and Evaluation of Antifungal Activity of 1-Butyl-3-hexyl-1H-imidazol-2(3H)-selenone by Surface-Enhanced Raman Spectroscopy.

In the present research work, a selenium N-heterocyclic carbene (Se-NHC) complex/adduct was synthesized and characterized by using different analytical methods including FT-IR, 1HNMR, and 13CNMR. The antifungal activity of the Se-NHC complex against Aspergillus flavus (A. flavus) fungus was investigated with disc diffusion assay. Moreover, the biochemical changes occurring in this fungus due to exposure of different concentrations of the in-house synthesized compound are characterized by surface-enhanced Raman spectroscopy (SERS) and are illustrated in the form of SERS spectral peaks. SERS analysis yields valuable information about the probable mechanisms responsible for the antifungal effects of the Se-NHC complex. As demonstrated by the SERS spectra, this Se-NHC complex caused denaturation and conformational changes in the proteins as well as decomposition of the fungal cell membrane. The SERS spectra were analyzed using two chemometric tools such as principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA). The fungal samples' SERS spectra were differentiated using PCA, while various groups of spectra were discriminated with ultrahigh sensitivity (98%), high specificity (99.7%), accuracy (100%), and area under the receiver operating characteristic curve (87%) using PLS-DA.

Enterotypes of the human gut mycobiome

BackgroundThe fungal component of the human gut microbiome, also known as the mycobiome, plays a vital role in intestinal ecology and human health. However, the overall structure of the gut mycobiome as well as the inter-individual variations in fungal composition remains largely unknown. In this study, we collected a total of 3363 fungal sequencing samples from 16 cohorts across three continents, including 572 newly profiled samples from China.ResultsWe identify and characterize four mycobiome enterotypes using ITS profiling of 3363 samples from 16 cohorts. These enterotypes exhibit stability across populations and geographical locations and significant correlation with bacterial enterotypes. Particularly, we notice that fungal enterotypes have a strong age preference, where the enterotype dominated by Candida (i.e., Can_type enterotype) is enriched in the elderly population and confers an increased risk of multiple diseases associated with a compromised intestinal barrier. In addition, bidirectional mediation analysis reveals that the fungi-contributed aerobic respiration pathway associated with the Can_type enterotype might mediate the association between the compromised intestinal barrier and aging.ConclusionsWe show that the human gut mycobiome has stable compositional patterns across individuals and significantly correlates with multiple host factors, such as diseases and host age.CJ96cDD7W-mAM3Q8ovNQ4eVideo

Prevalence of Candida Species Isolates in Patients with Urinary Tract Infection in Madinah, Saudi Arabia

INTRODUCTION: Urinary tract infections (UTIs) is considered one of the most prevalent infections that may lead to many renal complications or dysfunctions. They are responsible for almost 10% of all infections in Saudi Arabia, which makes them the second most common cause of emergency department admission. Bacteria are the most common pathogens associated with UTIs. Nevertheless, UTIs caused by fungi have also been reported. Among fungal infections, Candida spp. has been identified as the primary cause of UTI-related mycoses. 
 
 OBJECTIVE: To assess the prevalence of Candida species isolated from adult patients in Madinah, Saudi Arabia.
 
 MATERIALS & METHODS: A retrospective cross-sectional study was performed using data collected from patients who visited King Fahad General Hospital in Madinah, Saudi Arabia. A total of 16803 urine bacterial cultures data were collected from January 2019 to October 2021. 
 
 RESULTS: Among the 4281 positive urine cultures, 92% (n = 3937) showed bacterial growth, while 8% (n = 344) exhibited fungal growth. Female patients had a slightly higher percentage of fungal-positive cases (53%, n = 181) compared to males (47%, n = 163). Among different nationalities, Saudi citizens had the highest prevalence of positive fungal samples (64%, n = 220). Most positive cases (49%, n = 167) were isolated in 2019, with a decline in subsequent years. Non-Candida albicans spp. (86.3%, n = 297) were the most common fungal species, followed by Candida albicans (13.7%, n = 47).
 
 CONCLUSION: Despite considering bacterial UTIs to be more prevalent than UTI-related mycoses, it remains crucial to accurately identify the causative agent for proper diagnosis and treatment. UTI-related mycosis caused by non-Candida albicans spp. has significantly increased compared to Candida albicans. Thus, monitoring these trends over time can provide valuable insights for developing preventive strategies and optimizing treatment approaches.

Distribution of fungemia agents in five years and antifungal resistance.

Recent research has suggested that fungemia may demonstrate an epidemiologic shift in etiologic agents. This study focuses on the agents causing fungemia and antifungal resistance in a tertiary hospital. We evaluated all-age fungemia cases admitted to Balikesir Ataturk City Hospital in 2017-2021. Blood cultures (BC) were studied using BacT/Alert® 3D (bioMérieux, Marcyl'Etoile, France) and Render BC128 System (Render Biotech Co. Ltd., Shenzhen, China). On the data, we explored only the first fungal positive samples or the first isolates in different episodes of the same patients. Upon The Clinical and Laboratory Standards Institute (CLSI) disk diffusion guidelines, conventional methods and the Phoenix™ 100 System (Becton Dickinson, Franklin Lakes, NJ, USA) were utilized for antifungal susceptibility identifications. The findings showed that 325 (0.84%) of 38,682 BC sets were positive for fungal growth. Except for four cases (1.2%) [Saprochaete capitata (n = 2); Trichosporon asahii (n = 1), and Saccharomyces cerevisiae (n = 1)], all positive cases yielded Candida spp. (98.8%) growth. In these patients, the following Candida spp. were isolated: Candida albicans complex (n = 155; 47.7%), Candida parapsilosis complex (n = 127; 39.1%), Candida glabrata complex (n = 19; 5.85%), Candida tropicalis (n = 12; 3.7%), Candida kefyr (n = 5; 1.54%), Candida krusei (n = 2; 0.62%), and Candida guilliermondii complex (n = 1; 0.31%). We also realized that while none of the Candida spp. had echinocandin resistance, 8 C. parapsilosis complex isolates were resistant to fluconazole, and 17 C. parapsilosis complex and 2 C. tropicalis isolates were susceptible dose-dependent to fluconazole. In brief, antifungal resistance is more likely to restrict therapeutic options, albeit it is, fortunately, not prevalent in Turkey despite a few recent reports. Yet, a robust detection or management of antifungal resistance requires species-level identification and strict compliance with relevant management guidelines. Besides, challenges in research may be compensated with a national data set built with data from local laboratories.

Wood decay fungi show enhanced biodeterioration of low-density polyethylene in the absence of wood in culture media.

The involvement of microorganisms in low-density polyethylene (LDPE) degradation is widely studied across the globe. Even though soil, landfills, and garbage dumps are reported to be promising niches for such organisms, recently the involvement of wood decay fungi in polyethylene degradation is highlighted. In light of this, 50 fungal samples isolated from decaying hardwoods were assessed for their wood degradation ability and for their depolymerization enzymatic activities. For the LDPE deterioration assay, 22 fungal isolates having wood decay ability and de-polymerization enzymatic activities were selected. Fungal cultures with LDPE sheets (2 cm x 10 cm x 37.5 μm) were incubated in the presence and in the absence of wood as the carbon source (C) for 45 days. Degradation was measured by weight loss, changes in tensile properties, reduction in contact angle, changes of functional groups in Fourier-transform infrared spectroscopy, Scanning electron microscopic imaging, and CO2 evolution by strum test. Among the isolates incubated in the absence of wood, Phlebiopsis flavidoalba out-performed the other fungal species showing the highest percentage of weight reduction (23.68 ± 0.34%), and the lowest contact angle (64.28° ± 5.01). Biodegradation of LDPE by P. flavidoalba was further supported by 46.79 ± 0.67% of the mass loss, and 3.07 ± 0.13% of CO2 emission (mg/L) in the strum test. The most striking feature of the experiment was that all the isolates showed elevated degradation of LDPE in the absence of wood than that in the presence of wood. It is clear that in the absence of a preferred C source, wood decay fungi thrive to utilize any available C source (LDPE in this case) showing the metabolic adaptability of fungi to survive under stressful conditions. A potential mechanism for LDPE degradation is also proposed.

Hempcrete as a substrate for fungal growth under high humidity and variable temperature conditions

Bio-based alternatives for existing construction materials can be used to reduce the carbon footprint of the built environment. Hempcrete is one of these materials and is both an excellent hygric/thermal regulator and is carbon negative. However, this novel material is still incompletely researched, especially its fungal growth potential specifically within warm and humid environments. The incorporation of significant biological material within hempcrete can enable it to act as a microbial growth medium, with the corresponding potential for the release of bioaerosols. The aim of this research was thus to investigate the overall practicality of hempcrete implementation in a humid climate. To achieve this, the endogenous fungal genera on a sample of hempcrete were identified, fungal propagules aerosolized from a hempcrete sample enumerated, and a range of temperatures tested to determine their effect on fungi growth determined. Trials were performed to determine whether hempcrete can be effectively decontaminated with common materials to manage microbial growth. Under high humidity, fungal propagule emissions were high with low diversity, with potentially allergenic fungi detected. Disinfection of high fungal load hempcrete samples was able to reduce ∼94% of the fungal observations and reduce aerosolized counts to average background tropical fungal counts. The range of temperatures tested were not found to effect fungal growth, contrary to the consensus of the literature. Overall, these properties make hempcrete suited to humid areas, however, further research to investigate the potential effects of fungi on the material remains lacking.

Genetic diversity of culturable fungi associated with scleractinian corals in the Gulf of Thailand

Abstract Coral reefs are one of the most biodiverse marine ecosystems. Diverse microbes are associated with corals, including zooxanthellae, protists, prokaryotes, and viruses. This study aimed to investigate the genetic diversity of the culturable fungi associated with scleractinian corals in the Gulf of Thailand. Ribosomal DNA sequence analysis and morphological analysis of 130 fungal isolates revealed a high diversity of fungal phylotypes from three coral species: Pavona decussata, Pocillopora damicornis and Porites lutea. Ascomycota was the most abundant fungal phylum. The predominant orders included Eurotiales, Cladosporiales, Hypocreales, Mycosphaerellales, and Diaporthales. There were eight orders of the Basidiomycota dominated by Polyporales, Hymenochaetales, Agaricales, and Wallemiales. A principal component analysis was performed to compare abundance and correlation between the fungal communities, sampling locations and coral species. The results showed that the dissimilarity of fungal communities corresponded to geographic localities. On the other hand, there was a considerable overlap between the fungal communities of different coral species. The results from this research provide information on the diversity and ecology of the fungal communities associated with scleractinian corals in the Gulf of Thailand.

Ramularia leaf spot: PCR-based methods reveal widespread distribution of Ramulariopsis pseudoglycines and limited presence of R. gossypii in Brazil

Whilst Brazil is the fourth largest cotton producer globally, incidence of ramularia leaf spot (RLS) has decreased yield. In 2017–18 and 2018–19, ca. 300 fungal samples were collected throughout Brazil. Hyphal tip cultures were obtained for amplification of the RNA polymerase II (RPB2), 28S rRNA, the ribosomal DNA internal transcribed spacers (ITS), actin (ACT), elongation factor (EF1-α) and histone H3 (HIS3) genomic regions. Additionally, sequences of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were obtained by nanopore sequencing and the EF1-α region was selected as a marker for rapid recognition of Ramulariopsis species. Clade assignments based on the concatenated-sequence tree were identical to those in tree generated by RPB2-sequences, as well as in an RPB2 haplotype network and an ISSR (TGTC)4 dendrogram, in identification with species-specific primers and based on morphological comparisons. Out of 267 examined isolates, 252 were identified as Ramulariopsis pseudoglycines, indicating this species as the most widespread causal agent of cotton RLS in the Brazilian growing regions. Species-specific primers developed in the study that target the EF1-α gene provide an opportunity for extensive RLS sampling worldwide to study the distribution of Ramulariopsis species. Such data will aid breeders and plant pathologists in cotton disease resistance development and fungicide resistance avoidance.

Microbial Assemblages Associated with the Soil-Root Continuum of an Endangered Plant, Helianthemum songaricum Schrenk.

The microbial network of the soil-root continuum plays a key role in plant growth. To date, limited information is available about the microbial assemblages in the rhizosphere and endosphere of endangered plants. We suspect that unknown microorganisms in roots and soil play an important role in the survival strategies of endangered plants. To address this research gap, we investigated the diversity and composition of the microbial communities of the soil-root continuum of the endangered shrub Helianthemum songaricum and observed that the microbial communities and structures of the rhizosphere and endosphere samples were distinguishable. The dominant rhizosphere bacteria were Actinobacteria (36.98%) and Acidobacteria (18.15%), whereas most endophytes were Alphaproteobacteria (23.17%) as well as Actinobacteria (29.94%). The relative abundance of rhizosphere bacteria was higher than that in endosphere samples. Fungal rhizosphere and endophyte samples had approximately equal abundances of the Sordariomycetes (23%), while the Pezizomycetes were more abundant in the soil (31.95%) than in the roots (5.70%). The phylogenetic relationships of the abundances of microbes in root and soil samples also showed that the most abundant bacterial and fungal reads tended to be dominant in either the soil or root samples but not both. Additionally, Pearson correlation heatmap analysis showed that the diversity and composition of soil bacteria and fungi were closely related to pH, total nitrogen, total phosphorus, and organic matter, of which pH and organic matter were the main drivers. These results clarify the different patterns of microbial communities of the soil-root continuum, in support of the better conservation and utilization of endangered desert plants in Inner Mongolia. IMPORTANCE Microbial assemblages play significant roles in plant survival, health, and ecological services. The symbiosis between soil microorganisms and these plants and their interactions with soil factors are important features of the adaptation of desert plants to an arid and barren environment. Therefore, the profound study of the microbial diversity of rare desert plants can provide important data to support the protection and utilization of rare desert plants. Accordingly, in this study, high-throughput sequencing technology was applied to study the microbial diversity in plant roots and rhizosphere soils. We expect that research on the relationship between soil and root microbial diversity and the environment will improve the survival of endangered plants in this environment. In summary, this study is the first to study the microbial diversity and community structure of Helianthemum songaricum Schrenk and compare the diversity and composition of the root and soil microbiomes.

Antioxidant and antifungal efficacy of green synthesized silver nanoparticles

The use of herbs have gained momentum in the contemporary research, as they possess several biological activities. Green synthesised nanoparticles are extensively under exploration due to their multi-faceted roles. The present work details the comparative antioxidant potentials of the leaves and seeds extracts of the herbs Coriander sativum (coriander), Trachyspermum ammi (Ajwain), Trigonella foenum-graecum (menthi) and synthesis of silver nanoparticles. The antifungal effects of the synthesized nanoparticles against a pathogenic species of Aspergillus flavus and the mode of action was also analysed. Results of the investigation showed that the seed extract of Ajwain and leaf extract of coriander exhibited the highest antioxidant properties and were used as reducing agents for silver nanoparticles synthesis. The nanoparticles were characterized by UV-visible absorption spectrum, Scanning Electron Microscopy-Energy Dispersive Spectra (SEM-EDS) and Transmission Electron Microscopy (TEM). The silver nanoparticles showed enhanced antioxidant activity when compared to the extracts. The antifungal studies showed that both the nanoparticles were effective against A. flavus, while C. sativum silver nanoparticle (Cs-AgNP) exhibited the highest activity. The mechanism of antifungal effect showed that the nanoparticles exerted a membranolytic activity on the pathogenic A. flavus. SEM images showed highly wrinkled mycelia in nanoparticles-treated samples and membrane permeabilization was evident from the 1- N-phenyl napththyl amine assay in nanoparticle-treated fungal samples. The activity of a transmembrane enzyme calcium ATPase was also significantly decreased in treated samples confirming the membrane damage effects of nanoparticles. Among the nanoparticles, Cs-AgNP showed the maximum effect. The efficacy of these green synthesized nanoparticles provides insight into their use as effective antifungal agents.

Identification and susceptibility testing of oral candidiasis in advanced cancer patients

BackgroundPatients with advanced cancer are prone to develop different opportunistic oral infection due to anti-cancer treatment or the malignancies themselves. Studies of oral fungal samples show an increased prevalence of non-Candida albicans species in mixed oral infections with Candida albicans. Non-C. albicans and C. albicans are associated with varying degrees of resistance to azoles, which may have implications for treatment. This study aimed to assess the diversity and antifungal susceptibility of Candida species detected in the oral cavity.MethodsAn observational study with microbiological analysis was conducted. Clinical fungal isolates were collected from patients in a hospice unit in 2014–2016. Isolates were re-grown on chromID® Candida plates in 2020. Single colony of each species was re-cultivated and prepared for biochemical identification with a VITEK2® system and verified by gene sequencing. Etest was performed on RPMI agar, and the antifungals fluconazole, amphotericin B, anidulafungin and nystatin were applied.ResultsFifty-six isolates from 45 patients were identified. Seven different Candida species and one Saccharomyces species were detected. The results of biochemical identification were confirmed with sequencing analysis. Thirty-six patients had mono infection, and nine out of 45 patients had 2–3 different species detected. Of C. albicans strains, 39 out of 40 were susceptible to fluconazole. Two non-C. albicans species were resistant to fluconazole, one to amphotericin B and three to anidulafungin.ConclusionC. albicans was the predominant species, with a high susceptibility to antifungal agents. Different Candida species occur in both mono and mixed infections. Identification and susceptibility testing may therefore lead to more effective treatment and may prevent the development of resistance among patients with advanced cancer.Trail registrationThe study Oral Health in Advanced Cancer was registered at ClinicalTrials.gov (#NCT02067572) in 20/02/2014.

An autopsy case of disseminated Cunninghamella bertholletiae infection in an immunocompetent patient: a case report

BackgroundRecently, deaths due to mucormycosis in immunocompromised hosts have increased; however, the clinical and pathological features of mucormycosis are not fully understood, especially in view of the associated high mortality and rare incidence in immunocompetent patients.Case presentationWe have described a rare autopsy case of a 67-year-old Japanese man with chronic obstructive pulmonary disease who contracted mucormycosis. He had not been on any immunosuppressants, and his immune functions were intact. Since 3 days prior to admission to our hospital, he had experienced progressive dyspnea, productive cough, and fever. Chest computed tomography revealed pleural effusion in the left lower hemithorax and consolidation in the right lung field. Although he was administered with tazobactam-piperacillin hydrate (13.5 g/day), renal dysfunction occurred on the ninth disease day. Therefore, it was switched to cefepime (2 g/day). However, his general condition and lung-field abnormality worsened gradually. Cytological analysis of the sputum sample at admission mainly revealed sporangiophores and unicellular sporangioles, while repeated sputum culture yielded Cunninghamella species. Therefore, he was diagnosed with pulmonary mucormycosis. Liposomal amphotericin B (5 mg/kg/day) was initiated on the 28th disease day. However, chest radiography and electrocardiography detected cardiomegaly and atrial fibrillation, respectively, and he died on the 37th disease day. A postmortem examination revealed clusters of fungal hyphae within the arteries of the right pulmonary cavity wall, the subpericardial artery, intramyocardial capillary blood vessels, and the esophageal subserosa vein. Direct sequencing revealed that all fungal culture samples were positive for Cunninghamella bertholletiae.ConclusionsCunninghamella bertholletiae could rapidly progress from colonizing the bronchi to infecting the surrounding organs via vascular invasion even in immunocompetent patients.

Microbial communities of biochar amended forest soils in northwestern USA

Biochar is an environmental soil application tool used for climate change mitigation and carbon (C) sequestration that can be used in forest soils. Soil physical and chemical properties can be altered by biochar amendment, which can affect soil microbial communities. A limited number of forest soil studies have shown biochar can affect soil microbial communities, but few studies exist in temperate, northwestern USA forests. We investigated the effects of biochar amendment to soil in three western USA managed forest sites. Sites were amended with 0, 2.5, or 25 Mg ha−1 biochar, applied to the soil surface. DNA was extracted from soil samples collected from two depths, three to five years following amendment (dependent on site). Double-barcoded 16S rRNA (bacteria) and LSU rRNA (fungi) amplicons were sequenced on an Illumina MiSeq platform. Resulting sequencing data was analyzed for community richness, diversity, phyla relative abundance, and composition. For fungi, sequencing data represented six phyla. Biochar amendment did not affect fungal community measures, but site and sampling depth or their interaction did. Bacterial sequencing data represented 33 phyla. Biochar did not affect richness or diversity measures. However biochar did affect Bacteroidetes and Acidobacteria phyla relative abundance either alone, or when dependent on site or sampling depth. As with fungi, site and sampling depth or their interaction influenced richness, diversity, phyla relative abundance, and community composition. Results indicate that biochar amendment to northwestern USA forests is not detrimental to soil microbial community composition.

Dissemination of Fusarium Wilt of Banana in Mozambique Caused by Fusarium odoratissimum Tropical Race 4.

Fusarium wilt of banana (FWB) is a serious soil-borne fungal disease. In the previous century, FWB already destroyed Gros Michel-based banana cultures in Central America, and currently, the disease threatens all major banana-producing regions of the world. The causal agents of these epidemics, however, are diverse. Gros Michel was infected by a wide range of Fusarium species, the so-called Race 1 strains, whereas the contemporary Cavendish-based cultures are affected by Fusarium odoratissimum, colloquially called Tropical Race 4 (TR4). TR4 was reported in Mozambique on two commercial banana farms in 2013, but no incursions were found outside the farm boundaries in 2015, suggesting that the disease was under control. Here we report the presence of TR4 outside of these farm boundaries. We obtained fungal samples from 13 banana plants in smallholder and roadside plantings at various locations throughout northern Mozambique. These samples tested positive for TR4 by molecular diagnostics and in greenhouse pathogenicity assays. The results were confirmed with reisolations, thereby completing Koch's postulates. To study the diversity of TR4 isolates in Mozambique, we selected five samples for whole-genome sequencing. Comparison with a global collection of TR4 samples revealed very little genetic variation, indicating that the fungus is clonally spreading in Mozambique. Furthermore, isolates from Mozambique are clearly genetically separated from other geographic incursions, and thus we cannot trace the origin of TR4 in Mozambique. Nevertheless, our data demonstrates the dissemination of TR4 in Mozambique, underscoring the failure of disease management strategies. This threatens African banana production.

Quantitative Spectroscopic Characterization of Near-UV/visible E. coli (pYAC4), B. subtilis (PY79), and Green Bread Mold Fungus Fluorescence for Diagnostic Applications.

Ultraviolet (UV)-excited visible fluorescence is an attractive option for low-cost, low-complexity, rapid imaging of bacterial and fungal samples for imaging diagnostics in the biomedical community. While several studies have shown there is potential for identification of microbial samples, very little quantitative information is available in the literature for the purposes of diagnostic design. In this work, two non-pathogenic bacteria samples (E. coli pYAC4, and B. subtilis PY79) and a wild-cultivated green bread mold fungus sample are characterized spectroscopically for the purpose of diagnostic design. For each sample, fluorescence spectra excited with low-power near-UV continuous wave (CW) sources, and extinction and elastic scattering spectra are captured and compared. Absolute fluorescence intensity per cell excited at 340 nm is estimated from imaging measurements of aqueous samples. The results are used to estimate detection limits for a prototypical imaging experiment. It was found that fluorescence imaging is feasible for as few as 35 bacteria cells (or [Formula: see text]30 µm3 of bacteria) per pixel, and that the fluorescence intensity per unit volume is similar for the three samples tested here. A discussion and model of the mechanism of bacterial fluorescence in E. coli is provided.

Investigation of Virulence Factors in Microbial Organisms that Associated with Public Health Risk Isolates from Different Environmental Regions

Infectious diseases caused by infected tools in the environments are threaten to the safety and public health. Transmission sources of these infectious diseases are unknown, but it is thought that non-living materials called fomites, are the major source of acquired infections. Three hundred and one swabs were taken from different sources and cultured on blood agar to study heamolysis ability of isolated bacteria. In this study, MacConkey agar was used to isolate Gram-negative bacteria and Sabouraud agar (SDA) to isolate fungi. The biofilm formation test was done by Congo red plate assay. 41 (13.6%) bacterial isolates were obtained and (18.27%) of fungi were isolated on Sabouraud agar (SDA). Staphylococcus aureus was the more frequent bacterial species that isolated in this study. 29% of samples showed hemolysin activity on blood agar and 32%of the isolates were biofilm- producer. Results revealed that (7.9%) of Gram-negative bacteria harbored the fimH gene, (9%) harbored the icaA were Gram-positive and 6.3 % of fungal samples had HWP1 gene. Furthermore, (9.3%) from the total samples are bacterial samples harbored hla gene belong to Staphylococcus spp. Furthermore, (5.07%) of tested samples possessed hlyA gene were Gram-negative bacteria. We found in our study that infectious organisms can be transmitted from one individual to another by fomites responsible for acquired infection.

MnO2 nanoparticles/methylene blue-chitosan nanocomposite modified glassy carbon electrode (MnO2 NPs/MB–CS/GCE) as sensor for electrochemical determination of β-glucan in Morchella esculenta samples

The aim of this work was performed for design the electrochemical biosensor (βg-GOx/MnO2 NPs/MB-CS/GCE) determination of β-glucan in morel samples by modification of glassy carbon electrode (GCE) with electrodeposition of methylene blue-chitosan (MB-CS) nanocomposite and MnO2 nanoparticles (MnO2 NPs) and immobilization β-glucanase and glucose oxidase (βg-GOx), respectively. The study of the morphology of MnO2 NPs/MB–CS/GCE by FE-SEM indicated that the electrodeposited MnO2 NPs in the chitosan layer are wrapped by MB nanocubes. The electrochemical properties of β-glucan nonconductive enzyme film were successfully immobilized in β-glucan/MnO2 NPs/MB-CS/GCE, and the synergetic advantageous effect of MB-CS and MnO2 NPs enhanced the sensitivity and selectivity of the β-glucan biosensor. The linear range of 0– 10,400 ng/mL was obtained, and the sensitivity of 0.00441 µA/ng.mL−1 and the detection limit of 0.34 ng/mL were determined. The study on the applicability of the βg-GOx/MnO2 NPs/MB–CS/GCE as β-glucan sensor in morel samples revealed significant conformity between the results of amperometric and β-Glucan Assay Kit assays, and the obtained good recoveries between 90.00% and 97.50% with relative standard deviation less than 5.18% for amperometric analysis were acceptable, confirming the appropriate applicability of the βg-GOx/MnO2 NPs/MB–CS/GCE for determination of the β-glucan content in fungal samples.

Role of Next Generation Sequencing (NGS) in Plant Disease Management: A Review

A high throughput technique used to determine a part of the nucleotide sequence of an organism’s genome is called next generation sequencing (NGS). NGS has been Proven revolutionary in genomics. Clinical diagnostics, Plant diseases diagnostic and other aspects of medical are now made possible by sequencing. Techniques of NGS: there are different techniques of NGS which are being used in real life sciences i.e., Illumina sequencing, Pyrosequencing, Roche 454 sequencing and Ion torrent sequencing. All vintage methods like culturing in bacterial, fungal, and viral samples are being suppressed by next generation sequencing. The potential for random metagenomic sequencing of sick samples to find potential pathogens has surfaced with the development of next-generation high-throughput parallel sequencing technology. NGS enables highly efficient, rapid, low-cost DNA or RNA high-throughput sequencing of plant virus and viroids genomes, as well as specific small RNAs generated during infection. Although this technique is not so much familiar in the field of plant diseases. However, its widespread application in agronomic sciences will make it possible to create solutions to future food-related challenges that involve biotic stress.

Identification of Aspergillus Fumigatus using Molecular Techniques

Plant pathogenic fungi belong to the Kingdom Fungi comprising of over 100,000 registered species grouped into about 4,300 genera. Many of these fungi infect a variety of cereals particularly stored condition. A study was conducted to evaluate the occurrence of Plant pathogenic fungi associated with stored Sorghum vulgare grains. The fungal was subjected to the fungal load for exact identification using the molecular biology technique. The fungal isolates were isolated from the collected grains. Results revealed that Genus Aspergillus fumigatus was the most frequent isolate which are known to produce Aflatoxins. A combination of many different isolates was present on the grains. The molecular identification method using Polymerase Chain Reaction (PCR) helped in the identification of fungi at the species level with precision and in the least possible time. In conclusion, the isolated species were identified morphologically as Aspergillus fumigatus. The fungal samples were then identified through molecular techniques by DNA sequencing which was identified as Aspergillus fumigatus. The molecular techniques used in this study, has added great benefits to the process of distinguishing between similar species of fungi in comparison with the classical techniques. Furthermore, there is a need for having a National Level, Germplasm collection centre and detailed database of all the naturally occurring post-harvest Plant pathogenic fungi for easy intervention and timely overcoming of situations in case of any emergency.

Optical sectioning of label-free samples using standard bright-field microscopy.

The three-dimensional (3D) images produced by the traditional bright-field (BF) light microscope are ill-defined and therefore require reconstruction. Current alternatives to perform 3D BF imaging require dedicated instrumentation or extensive computer modeling. We report optical sectioning in bright-field microscopy (OSBM), a direct method for 3D imaging of label-free samples, where the BF inverse-imaging problem is solved in real space by using an elemental combination of hardware and software. Our method consists in acquiring z-stack images of minimally treated samples under Köhler illumination in the coherent regime, followed by a digital image processing pipeline designed to localize the source of axial intensity gradients that correspond to the z-locations of scatterers within the sample. We validate OSBM by visualizing fungal, animal tissue, and plant samples and comparing with light sheet fluorescence microscopy imaging, finding excellent match for sparse spatial distributions. Our work demonstrates how the standard microscope can be used as an effective 3D imaging device.