Fungi-derived agriculturally important nanoparticles and their application in crop stress management - Prospects and environmental risks.

Nanotechnology has a wide range of agricultural applications, with emphasize on the development of novel nano-agrochemicals such as, nano-fertilizer and nano-pesticides. It has a significant impact on sustainable agriculture by increasing agricultural productivity, while reducing the use of inorganic fertilizers, pesticides, and herbicides. Nano-coating delivery methods for agrochemicals have improved agrochemical effectiveness, safety, and consistency. Biosynthesis of nanoparticles (NPs) has recently been recognized as an effective tool, contrary to chemically derived NPs, for plant abiotic and biotic stress control, and crop improvement. In this regard, fungi have tremendous scope and importance for producing biogenic NPs of various sizes, shapes, and characteristics. Fungi are potential candidates for synthesis of biogenic NPs due to their enhanced bioavailability, biological activity, and higher metal tolerance. However, their biomimetic properties and high capacity for dispersion in soil, water environments, and foods may have negative environmental consequences. Furthermore, their bioaccumulation raises significant concerns about the novel properties of nanomaterials potentially causing adverse biological effects, including toxicity. This review provides a concise outline of the growing role of fungal-mediated metal NPs synthesis, its potential applications in crop field, and associated issues of nano-pollution in soil and its future implications.

Efficacy of a rhamnolipid biosurfactant to inhibit Trichophyton rubrum in vitro and in a mice model of dermatophytosis.

Dermatophytic infections caused by Trichophyton rubrum are the most prevalent superficial mycoses worldwide. The present study was aimed at evaluating the anti-dermatophytic effect of a rhamnolipid biosurfactant (RL) produced by Pseudomonas aeruginosa SS14 (GenBank Accession no. KC866140) against T. rubrum in experimentally induced dermatophytosis in mice models. The purified RL could effectively suppress spore germination and hyphal proliferation of T. rubrum at 500 µg/mL. Ultramicroscopic observations involving SEM and AFM studies revealed severely altered hyphal morphology in the RL-treated mycelia. The membrane disruptive effect of RL was measured by the uptake of propidium iodide (PI) as visualized by CLSM. The absorbance of the RL-treated cell suspension at 260 nm showed concentration-dependant exudation of nucleic acid due to loss of cell membrane integrity. On evaluation of the therapeutic efficacy of RL on experimentally induced cutaneous dermatophytosis in mice models, we observed that the RL, when applied topically at a concentration of 500 µg/mL, was effective in completely curing dermatophytosis at the end of 21-day treatment period. The results were statistically similar to those obtained using the standard drug terbinafine as control. Apart from macroscopic observation, the results were confirmed by culture of skin scrapings and histopathological examination. The results are suggestive of the effectiveness of RL in control of dermatophytosis caused by T. rubrum.

Production, characterization, and antifungal activity of a biosurfactant produced by Rhodotorula babjevae YS3.

BACKGROUND: Sophorolipids are one of the most promising glycolipid biosurfactants and have been successfully employed in bioremediation and various other industrial sectors. They have also been described to exhibit antimicrobial activity against different bacterial species. Nevertheless, previous literature pertaining to the antifungal activity of sophorolipids are limited indicating the need for further research to explore novel strains with wide antimicrobial activity. A novel yeast strain, Rhodotorula babjevae YS3, was recently isolated from an agricultural field in Assam, Northeast India. This study was primarily emphasized at the characterization and subsequent evaluation of antifungal activity of the sophorolipid biosurfactant produced by R. babjevae YS3. RESULTS: The growth kinetics and biosurfactant production by R. babjevae YS3 was evaluated by cultivation in Bushnell-Haas medium containing glucose (10% w/v) as the sole carbon source. A reduction in the surface tension of the culture medium from 70 to 32.6 mN/m was observed after 24 h. The yield of crude biosurfactant was recorded to be 19.0 g/l which might further increase after optimization of the growth parameters. The biosurfactant was characterized to be a heterogeneous sophorolipid (SL) with both lactonic and acidic forms after TLC, FTIR and LC-MS analyses. The SL exhibited excellent oil spreading and emulsifying activity against crude oil at 38.46 mm2 and 100% respectively. The CMC was observed to be 130 mg/l. The stability of the SL was evaluated over a wide range of pH (2-10), salinity (2-10% NaCl) and temperature (at 120 °C for time intervals of 30 up to 120 min). The SL was found to retain surface-active properties under the extreme conditions. Additionally, the SL exhibited promising antifungal activity against a considerably broad group of pathogenic fungi viz. Colletotrichum gloeosporioides, Fusarium verticilliodes, Fusarium oxysporum f. sp. pisi, Corynespora cassiicola, and Trichophyton rubrum. CONCLUSIONS: The study reports, for the first time, the biosurfactant producing ability of R. babjevae, a relatively lesser studied yeast. The persistent surface active properties of the sophorolipid in extreme conditions advocates its applicability in diverse environmental and industrial sectors. Further, antifungal activities against plant and human pathogens opens up possibilities for development of efficient and eco-friendly antifungal agents with agricultural and biomedical applications.

Transmission of African swine fever virus to the wild boars of Northeast India.

BACKGROUND: India recorded the first outbreak of African swine fever (ASF) in North-eastern region (NER) in the year 2020. AIM: The current study was undertaken to investigate the transmission of African swine fever virus (ASFV) in the wild boars of Northeast India, particularly of Assam. MATERIAL AND METHODS: ASF suspected mortal tissue remains and blood samples of wild boars collected from different locations of Assam were screened for molecular detection of swine viruses which includes Classical swine fever virus, Porcine Circovirus 2, Porcine reproductive and respiratory syndrome virus and ASFV. RESULTS: One sample each from Manas and Nameri National Parks was detected positive for ASFV. Besides this, one of the samples was positive for CSFV and one of the ASFV positive samples was also positive for PCV2. Several striking gross and microscopic alterations were noticed in different organs of ASFV infected animals. Sequencing and phylogenetic analysis of B646L gene confirmed the presence of ASFV genotype-II in wild boars. Circulation of similar genotype in domestic pigs of NER in the contemporary period as well as locations near to the aforementioned national parks indicates the transmission of ASFV from domestic to wild boars. CLINICAL RELEVANCE: The detection of ASFV in the wild boars of Assam is alarming as it is an impending threat to pig population and other endangered species (particularly Pygmy hog), making it increasingly daunting to control the disease. CONCLUSION: Chances are high for ASFV to become endemic in Assam region if stringent measures are not taken at proper time.

Draft Genome Sequence Analysis of the Genotype II African Swine Fever Virus from India.

African swine fever virus (ASFV) entered the northeastern (NE) part of India early in 2020, causing huge economic loss to the piggery sector. Here, we are presenting a brief report on the draft genome sequence of an ASFV strain ABTCVSCK_ASF007 from Assam state of NE India belonging to genotype II.

Selenite bioreduction and biosynthesis of selenium nanoparticles by Bacillus paramycoides SP3 isolated from coal mine overburden leachate.

A native strain of Bacillus paramycoides isolated from the leachate of coal mine overburden rocks was investigated for its potential to produce selenium nanoparticles (SeNPs) by biogenic reduction of selenite, one of the most toxic forms of selenium. 16S rDNA sequencing was used to identify the bacterial strain (SP3). The SeNPs were characterized using spectroscopic (UV-Vis absorbance, dynamic light scattering, X-ray diffraction, and Raman), surface charge measurement (zeta potential), and ultramicroscopic (FESEM, EDX, FETEM) analyses. SP3 exhibited extremely high selenite tolerance (1000 mM) and reduced 10 mM selenite under 72 h to produce spherical monodisperse SeNPs with an average size of 149.1 ± 29 nm. FTIR analyses indicated exopolysaccharides coating the surface of SeNPs, which imparted a charge of -29.9 mV (zeta potential). The XRD and Raman spectra revealed the SeNPs to be amorphous. Furthermore, biochemical assays and microscopic studies suggest that selenite was reduced by membrane reductases. This study reports, for the first time, the reduction of selenite and biosynthesis of SeNPs by B. paramycoides, a recently discovered bacterium. The results suggest that B. paramycoides SP3 could be exploited for eco-friendly removal of selenite from contaminated sites with the concomitant biosynthesis of SeNPs.

Rhamnolipid exhibits anti-biofilm activity against the dermatophytic fungi Trichophyton rubrum and Trichophyton mentagrophytes.

Dermatophytes are responsible for a majority of fungal infections in humans and other vertebrates, causing dermatophytosis. Treatment failures are often associated with biofilm formation, making dermatophytes resistant to antifungals. In this study, effects of a rhamnolipid (RL-SS14) produced by Pseudomonas aeruginosa SS14 on planktonic cells of Trichophyton rubrum and Trichophyton mentagrophytes, their biofilm formation, and disruption of mature biofilms were assessed. The composition of RL-SS14 was analysed using FTIR, HPLC-ESI-MS, and GC-MS. Minimum inhibitory concentrations against the planktonic forms of T. rubrum and T. mentagrophytes were 0.5 mg/mL and 0.125 mg/mL, respectively. Crystal-violet (biofilm biomass) and safranin (extracellular matrix) staining revealed that RL-SS14 significantly inhibited biofilm formation and also reduced preformed biofilms in a dose-dependent manner. Microscopic visualization of treated biofilms via SEM, AFM, and CLSM revealed marked morphological damage, cell death, and reduced extracellular matrix. The results indicate the potential of RL-SS14 as an anti-biofilm agent against dermatophytes.

Sophorolipid Biosurfactant Can Control Cutaneous Dermatophytosis Caused by Trichophyton mentagrophytes.

Trichophyton mentagrophytes, a zoophilic species, is one of the most frequently isolated dermatophytes in many parts of the world. This study investigated the efficacy of a sophorolipid (SL-YS3) produced by Rhodotorula babjevae YS3 against dermatophytosis caused by T. mentagrophytes. SL-YS3 was characterized by gas chromatography-mass spectrometry (GC-MS) and ultra-performance liquid chromatography, coupled with electrospray mass spectrometry (UPLC-ESI-MS). SL-YS3 comprised of six different fatty acids as the hydrophobic components of constituent congeners and sophorose as the hydrophilic component. Inhibitory effects of purified SL-YS3 against hyphal growth was found to be 85% at a 2 mg ml-1 concentration, and MIC was 1 mg ml-1. Microscopic examination with scanning electron microscopy (SEM), atomic force microscopy, and confocal laser scanning microscopy (CLSM) revealed that SL-YS3 exerts its effect by disrupting cell membrane integrity causing cell death. SL-YS3 was also effective in reducing the biofilms formed by T. mentagrophytes, which was observed spectrophotometrically with crystal-violet staining and further validated with SEM and CLSM studies of treated biofilms. In vivo studies in a mouse model of cutaneous dermatophytosis involving macroscopic observations, percent culture recovery from skin samples, and histopathological studies showed that SL-YS3 could effectively cure the infected mice after 21 days of topical treatment. Terbinafine (TRB) was used as a standard drug in the experiments. We demonstrate, for the first time, the antidermatophytic activity of a sophorolipid biosurfactant. The findings are suggestive that SL-YS3 can be formulated as a novel antifungal compound to treat cutaneous mycoses caused by T. mentagrophytes.

Rice based distillers dried grains with solubles as a low cost substrate for the production of a novel rhamnolipid biosurfactant having anti-biofilm activity against Candida tropicalis.

In this study, rhamnolipid (RL) production by Pseudomonas aeruginosa SS14 utilizing rice based Distillers Dried Grains with Solubles (rDDGS) as the sole carbon source was evaluated and the production parameters were optimized using response surface methodology. Highest RL (RL-rDDGS) yield was 14.87 g/L in a culture medium containing 12% (w/v) rDDGS and 11% (v/v) inoculum concentration after 48 h of fermentation at 35 °C. RL-rDDGS was produced as a mixture of mono and di-RL congeners with four novel homologues Rha-C18:2, Rha-C19, Rha-C9, and Rha-Rha-C19. The RL reduced the surface tension of water to 34.8 mN/m at a critical micelle concentration (CMC) value of 100 mg/L, exhibited high stability at a wide range of pH (6-12), heating time (0-120 min), and salinity (2-12% NaCl). Furthermore, RL-rDDGS demonstrated appreciable biofilm disruptive property against Candida tropicalis. This is the first report on the usage of rDDGS for sustainable and low cost production of RL.

Nanoencapsulated betulinic acid analogue distinctively improves colorectal carcinoma in vitro and in vivo.

Betulinic acid, a plant secondary metabolite, has gained significant attention due to its antiproliferative activity over a range of cancer cells. A promising betulinic acid analogue (2c) with better therapeutic efficacy than parent molecule to colon carcinoma cells has been reported. Despite impressive biological applications, low aqueous solubility and bioavailability create difficulties for its therapeutic applications. To overcome these lacunae and make it as a promising drug candidate we have encapsulated the lead betulinic acid derivative (2c) in a polymeric nanocarrier system (2c-NP) and evaluated its in vitro and in vivo therapeutic efficacy. Apoptosis that induces in vitro antiproliferative activity was significantly increased by 2c-NP compared to free-drug (2c), as assured by MTT assay, Annexin V positivity, JC1 analysis and cell cycle study. The therapeutic potential measured in vitro and in vivo reflects ability of 2c-NP as an effective therapeutic agent for treatment of colon carcinoma and future translation to clinical trials.