Alternaria alternata (SDHY01/02), a fungus associated with Lamellodysidea herbacea: its anticancer potential and responsible constituent(s).

One of the biggest global causes of death is cancer. The side effects of currently available therapies have triggered the search for new drugs. The marine environment, with its vast biodiversity, including sponges, is a rich source of natural products with immense pharmaceutical potential. The aim of the study was to analyze the microbes associated with the marine sponge, Lamellodysidea herbacea, and explore them as resources for anticancer ability. This study includes the isolation of fungi from L. herbacea, and their evaluation for cytotoxic potential against human cancer cell lines such as A-549 (lung), HCT-116 (colorectal carcinoma), HT-1080 (Fibrosarcoma), and PC-3 (prostate) using MTT assay. This revealed that fifteen extracts showed significant anticancer ability (IC50 ≤ 20 µg/mL), at least against one of the cell lines. Three extracts, SPG12, SPG19, and SDHY 01/02, were found significant in terms of anticancer activity, at least against three to four cell lines (IC50 values ≤ 20 µg/mL). The fungus SDHY01/02 was identified by sequencing the internal transcribed spacer (ITS) region as Alternaria alternata. Its extract showed IC50 values < 10 µg/mL against all the tested cell lines and was further analysed through light and fluorescence microscopy. The extract of SDHY01/02 was active (lowest IC50 4.27 µg/mL) against A549 cells in a dose-dependent manner and caused apoptotic cell death. Further, the extract was fractionated and analyzed the constituents by GC-MS (Gas Chromatography-Mass Spectrometry). Di-ethyl ether fraction revealed the constituents (having anticancer activity) such pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-3-(2-methyl propyl); 4,5,6,7-tetrahydro-benzo[C]rhiophene-1-carboxylic acid cyclopropylamide; 17-pentatriacontene; 9,12-octadecadienoic acid (Z, Z)-, methyl ester; while DCM fraction contained Oleic acid, eicosyl ester. This is the first report of A. alternata with anticancer potential that has been isolated from the sponge L. herbacea, as far as we are aware.This A. alternata can be exploited to get anticancer molecule(s) in the future.

A combinatorial approach of Monarda citriodora essential oil (MEO) and linalool vapors to control fruit rot of Citrus limon caused by a new pathogen, Aspergillus foetidus, and its underlying mode of action.

AIMS: Citrus limon (lemon) is a widely cultivated citrus fruit. Significant postharvest losses due to fungi plague its production. Environmental and human health hazards have made the application of synthetic fungicides unsuitable. Despite the previous reports of antifungal activities of essential oil (EO) vapors, their synergistic combinations are understudied. Synergistic vapor combinations are advantageous due to less concentration of active components. This study aimed to isolate and identify postharvest fungal pathogens lemon and to evaluate the antifungal effects of synergistic Monarda citriodora EO (MEO)-constituent vapor combinations in vivo and in vitro. METHODS AND RESULTS: Postharvest fungal pathogens of lemon (C. limon) were isolated from various infected samples. The most pathogenic isolate was identified through morphology and its ITS-based rRNA gene sequencing as Aspergillus foetidus (O4). This is the first report of A. foetidus as a postharvest pathogen of lemon. The minimum fungicidal concentrations (MFCs) of MEO vapors treatment against O4 were 1346.15 µL/L air. For carvacrol, hexanal, and linalool, MFC was same (96.16 µL/L air). Checkerboard assays demonstrated that 1/4 MFC of MEO (336.54 µL/L air) and 1/4 MFC of linalool (24.04 µL/L air) (M + L) were synergistic against O4. M + L vapors reduced the O4 growth on lemons during storage by 64% ± 1.50% and preserved their quality (low weight loss %, unchanged pH, increased ascorbic acid content). Propidium iodide staining, ergosterol content analysis, calcofluor white staining and chitin content analysis revealed the integrity loss of the O4 plasma membrane and cell wall. 2',7'-Dichlorofluorescin diacetate staining revealed accumulation of intracellular reactive oxygen species (ROS), and scanning electron microscopy (SEM) analysis exposed the M + L treated mycelia with malformations. CONCLUSIONS: M + L vapors offer protection for lemons from A. foetidus and preserve their quality during storage.

Management of postharvest blue mould of apple caused by Penicillium fuscoglaucum using a gel formulation containing Monarda citriodora essential oil and linalool.

AIMS: This study aimed to develop essential oil-containing biopolymer-based vapor gel formulations for the postharvest management of blue mould of apples. Apples are one of the widely cultivated fruits. They are susceptible to a wide range of fungus leading to high losses in overall production. Many research articles have highlighted the applications of essential oil-biopolymer coatings in the postharvest storage of fruits. However, no studies have yet tried to explore the potential of a vapour gel formulation for postharvest applications. METHODS AND RESULTS: Contaminated apples were collected from the local market. The causative fungus was isolated and identified. Minimum fungicidal concentrations of Monarda citriodora essential oil (MEO) and hexanal/linalool in the vapour phase were determined in vitro. In vitro and in vivo, checkerboard assays were used to demonstrate the synergistic activity between MEO and hexanal/linalool vapours against the isolated pathogen. MEO and linalool (M + L) combinatorial treatment indicated synergy in vivo and in vitro. In vivo treatment of M + L on apples by direct fumigation showed phytotoxicity. Gel formulations (carrageenan-guar gum, carbopol gel, and soft gel) were developed and evaluated to address phytotoxicity. M + L combination synergistically remediated the phytotoxicity in both carbopol (FICI = 0.625) and soft gels (FICI = 0.5625). The physicochemical parameters (pH, weight loss, TSS, and ascorbic acid (AsAC) of the treated apples were analysed. Reduction in weight loss and increase in AsAC were observed for treated fruits when compared to control, and no change in pH and TSS was observed. CONCLUSIONS: Biopolymer vapour gel formulations containing M + L vapours effectively protect apples from the postharvest blue mould for extended storage.

Myxobacteria from animal dung pellets collected from northwestern Himalayas: A new source of di-isobutyl phthalate.

Myxobacteria have emerged as a rich manufacturer of a wide array of natural products captivating both the academic and drug discovery communities. Attempts to unearth novel bioactive, myxobacteria from unexploited habitats are far from exhaustion. This study reports the isolation of myxobacteria from dung pellets collected from various regions of northwestern Himalayas. The isolated myxobacteria were functionally characterized to evaluate their bioactive capability. Of all the isolates, ST/P/71 exhibited broad range activities such as anticancer against all the four human cancer cell lines with IC50 in range of 2.03-9.65 µg/ml, antimicrobial against all the tested human pathogens, also exhibiting biofilm inhibition with MBIC50 at 10.4 µg/ml against Salmonella typhimurium. Consequently, ST/P/71 was chosen for fermentation and isolation of bioactive secondary metabolite through semi-preparative HPLC. It yielded compound 1, characterized as di-isobutyl phthalate (DiBP) based on nuclear magnetic resonance (NMR) and mass data. DiBP exhibited promising cytotoxic activity against the lung cancer cell line (A549) at an IC50 values 3.09 µg/ml and biofilm inhibition activity against Bacillus subtilis and Salmonella typhimurium with MBIC50 2.703 and 9.263 µg/ml, respectively. ST/P/71 was identified as Myxococcus fulvus. Thus, M. fulvus ST/P/71 isolated from northwestern Himalayas is a new source of DiBP.

Investigation of α/γ hybrid peptide self-assembled structures with antimicrobial and antibiofilm properties.

The present work describes the synthesis and characterization of α/γ hybrid peptides, Boc-Phe-γ4 -Phe-Val-OMe, P1; Boc-Ala-γ4 -Phe-Val-OMe, P2; and Boc-Leu-γ4 -Phe-Val-OMe, P3 together with the formation of self-assembled structures formed by these hybrid peptides in dimethyl sulfoxide (DMSO)/water (1:1). The self-assembled structures were characterized by infrared (IR) spectroscopy, circular dichroism (CD), and scanning electron microscopy (SEM). Further, α/γ hybrid peptide self-assembled structures were evaluated for antibacterial properties. Among all, the self-assembled peptide P1 exhibited the antimicrobial activity against Escherichia coli and Klebsiella pneumoniae, while self-assembled peptide P3 inhibited the biofilms of Salmonella typhimurium and Pseudomonas aeruginosa. In this study, we have shown the significance of self-assembled structures formed from completely hydrophobic α/γ hybrid peptides in exploring the antibacterial properties together with biofilm inhibition.

Trichoderma lixii (IIIM-B4), an endophyte of Bacopa monnieri L. producing peptaibols.

BACKGROUND: Exploration of microbes isolated from north western Himalayas for bioactive natural products. RESULTS: A strain of Trichoderma lixii (IIIM-B4) was isolated from Bacopa monnieri L. The ITS based rDNA gene sequence of strain IIIM-B4 displayed 99% sequence similarity with different Trichoderma harzianum species complex. The highest score was displayed for Hypocrea lixii strain FJ462763 followed by H. nigricans strain NBRC31285, Trichoderma lixii strain CBS 110080, T. afroharzianum strain CBS124620 and Trichoderma guizhouense BPI:GJS 08135 respectively. Position of T. lixii (IIIM-B4) in phylogenetic tree suggested separate identity of the strain. Microbial dynamics of T. lixii (IIIM-B4) was investigated for small peptides. Medium to long chain length peptaibols of 11 residue (Group A), 14 residue (Group B) and 17 residue (Group C) were identified using Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometer. Optimization is undeniably a desideratum for maximized production of desirable metabolites from microbial strain. Here optimization studies were carried out on T. lixii (IIIM-B4) using different growth media through Intact Cell Mass Spectrometry (ICMS). A multifold increase was obtained in production of 11 residue peptaibols using rose bengal medium. Out of these, one of them named as Tribacopin AV was isolated and sequenced through mass studied. It was found novel as having unique sequence Ac-Gly-Leu-Leu-Leu-Ala-Leu-Pro-Leu-Aib-Val-Gln-OH. It was found to have antifungal activity against Candida albicans (25 µg/mL MIC). CONCLUSION: In this study, we isolated a strain of T. lixii (IIIM-B4) producing medium and long chain peptaibols. One of them named as Tribacopin AV was found novel as having unique sequence Ac-Gly-Leu-Leu-Leu-Ala-Leu-Pro-Leu-Aib-Val-Gln-OH, which had antifungal properties.

An endophytic Fusarium sp. isolated from Monarda citriodora produces the industrially important plant-like volatile organic compound hexanal.

An endophytic fungus, MC_25L, has been isolated from the leaves of MonardacitriodoraCerv. ex Lag., a medicinal and aromatic herb from the northwestern Himalayas. It produces a fruity fragrance while growing on potato dextrose agar, suggesting that it is producing volatile organic compounds (VOCs). The endophyte inhibited the growth of plant pathogens such asSclerotiniasp. and Aspergillusflavus by virtue of VOCs. Identification of MC_25L based on morphological and microscopic features, as well as ITS-based rDNA sequence analysis, revealed that it is a Fusariumsp. GC-MS analysis revealed that this endophyte produces a unique array of VOCs, in particular hexanal, p-fluoroanisole, pentafluoropropionic acid 2-ethylhexyl, (5E)-5-ethyl-2-methyl-5-hepten-3-one, 2-butyl-2-hexanol, (7E)-2-methyl-7-hexadecene and acoradiene. Three major compounds were hexanal, (5E)-5-ethyl-2-methyl-5-hepten-3-one and acoradiene, and they account for around 84.57 % of the total VOCs. Moreover, of interest was the presence of hexanal, which has applications in the food and cosmetic industries, as well as in mycofumigation. This is the first report of a fungal endophyte producing the industrially important plant-like VOC hexanal. Hexanal is also active biologically. Thus this study indicates that Fusariumsp. (MC_25L) is a potential candidate for the up-scaling of hexanal.

Diversity, Phylogeny, anticancer and antimicrobial potential of fungal endophytes associated with Monarda citriodora L.

BACKGROUND: Present study focuses on diversity and distribution analysis of endophytic fungi associated with different tissues of the Monarda citriodora Cerv. ex Lag. (Lamiaceae/Labiatae). Anticancer and antimicrobial potential of isolated endophytes have also been investigated. RESULTS: A total of twenty eight fungal endophytes belonging to 11 different genera were isolated from this plant. All the endophytic fungi belonged to the Ascomycota phylum. The leaves were immensely rich in fungal species, while roots showed the highest tissue specific fungal dominance. Out of 28 fungal species, 72% endophytic extracts were found cytotoxic against one or more human cancer cell lines. The most prominent anticancer activity (IC50 value <10 µg/mL) was shown by MC-14 L (Fusarium oxysporum), MC-14 F (F. oxysporum), MC-18 L (Aspergillus fumigatus), MC-24 L (Cladosporium tenuissimum), MC-25 L (Fusarium sp.), MC-26 F (F. oxysporum) extracts. 75% of the extracts showed antimicrobial activities in agar disc-diffusion assay and 27% in the tube dilution method (MIC <100 µg/mL) respectively against the tested pathogens. Extracts of MC-14 L (F. oxysporum) and MC-18 L (A. fumigatus) displayed broad spectrum antimicrobial activity. CONCLUSIONS: These results indicated that M. citriodora harbors a rich fungal endophytic community with anticancer and antimicrobial activities. The isolated endophyte MC-24 L (C. tenuissimum) has the potential to be a source of novel cytotoxic/antimicrobial compounds. This is the first report of diversity of fungal endophytes isolated from M. citriodora.

Endophytic fungi associated with Monarda citriodora, an aromatic and medicinal plant and their biocontrol potential.

CONTEXT: The Food and Agriculture Organization has estimated that every year considerable losses of the food crops occur due to plant diseases. Although fungicides are extensively used for management of plant diseases, they are expensive and hazardous to the environment and human health. Alternatively, biological control is the safe way to overcome the effects of plant diseases and to sustain agriculture. Since Monarda citriodora Cerv. ex Lag. (Lamiaceae/Labiatae) is known for its antifungal properties, it was chosen for the study. OBJECTIVE: The isolation of endophytic fungi from M. citriodora and assessing their biocontrol potential. MATERIAL AND METHODS: The isolated endophytes were characterized using ITS-5.8 S rDNA sequencing. Their biocontrol potential was assessed using different antagonistic assays against major plant pathogens. RESULTS: Twenty-eight endophytes representing 11 genera were isolated, of which, around 82% endophytes showed biocontrol potential against plant pathogens. MC-2 L (Fusarium oxysporum), MC-14 F (F. oxysporum), MC-22 F (F. oxysporum) and MC-25 F (F. redolens) displayed significant antagonistic activity against all the tested pathogens. Interestingly, MC-10 L (Muscodor yucatanensis) completely inhibited the growth of Sclerotinia sp., Colletotrichum capsici, Aspergillus flavus and A. fumigatus in dual culture assay, whereas MC-8 L (A. oryzae) and MC-9 L (Penicillium commune) completely inhibited the growth of the Sclerotinia sp. in fumigation assay. CONCLUSIONS: Endophytes MC-2 L, MC-14 F, MC-22 F and MC-25 F could effectively be used to control broad range of phytopathogens, while MC-10 L, MC-8 L and MC-9 L could be used to control specific pathogens. Secondly, endophytes showing varying degrees of antagonism in different assays represented the chemo-diversity not only as promising biocontrol agents but also as a resource of defensive and bioactive metabolites.

Heterologous Production of Cyanobacterial Mycosporine-Like Amino Acids Mycosporine-Ornithine and Mycosporine-Lysine in Escherichia coli.

Mycosporine-like amino acids (MAAs) are an important class of secondary metabolites known for their protection against UV radiation and other stress factors. Cyanobacteria produce a variety of MAAs, including shinorine, the active ingredient in many sunscreen creams. Bioinformatic analysis of the genome of the soil-dwelling cyanobacterium Cylindrospermum stagnale PCC 7417 revealed a new gene cluster with homology to MAA synthase from Nostoc punctiforme This newly identified gene cluster is unusual because it has five biosynthesis genes (mylA to mylE), compared to the four found in other MAA gene clusters. Heterologous expression of mylA to mylE in Escherichia coli resulted in the production of mycosporine-lysine and the novel compound mycosporine-ornithine. To our knowledge, this is the first time these compounds have been heterologously produced in E. coli and structurally characterized via direct spectral guidance. This study offers insight into the diversity, biosynthesis, and structure of cyanobacterial MAAs and highlights their amenability to heterologous production methods. IMPORTANCE: Mycosporine-like amino acids (MAAs) are significant from an environmental microbiological perspective as they offer microbes protection against a variety of stress factors, including UV radiation. The heterologous expression of MAAs in E. coli is also significant from a biotechnological perspective as MAAs are the active ingredient in next-generation sunscreens.

Immunosuppressive potential of Botryosphaeria dothidea, an endophyte isolated from Kigelia africana.

CONTEXT: For years, natural products from microbes have been used as drugs. Endophytes are the most important fungi that produce many novel metabolites for potential use in pharmacology and agriculture. OBJECTIVE: The objective of the present study was to explore new endophytes for novel natural products. MATERIALS AND METHODS: An endophyte BAK-I was isolated from the bark of Kigelia africana (Lam.) Beneth (Bignoniaceae). BAK-I was characterized morphologically and on the basis of ITS-5.8S rDNA sequences. BAK-I was fermented to yield an extract, which was evaluated for its anticancer, antimicrobial, and immunomodulatory activities, using MTT, agar well-diffusion, tube dilution method, lymphocyte proliferation, and pro-inflammatory cytokines (TNF-α) (by macrophages) evaluation assays. For lymphocyte proliferation and pro-inflammatory cytokines studies, four concentrations were evaluated 10, 30, 100, and 1000 µg/mL and the experiments were conducted for 72 and 48 h, respectively. RESULTS AND DISCUSSION: The BAK-I showed pink cottony growth. SEM studies showed smooth fusoid-oblong conidia with a truncated base. Furthermore, ITS-5.8S rDNA sequence showed 99% homology with the Botryosphaeria dothidea strain suggesting that the endophyte is a strain of the genus Botryosphaeria. Less than 50% growth inhibition of SF295, Lung A-549, and THP-1 cancer cell lines after treatment with BAK-I extract suggested that it did not have significant cytotoxic potential, whereas it is bactericidal for Gram-positive pathogens MRSA and VRE with MIC value 200 and 250 µg/mL, respectively. To elucidate its immunomodulation potential, splenocyte proliferation studies showed that BAK-1 suppressed the T cell proliferation by 50%. TNF-α evaluation studies also showed that the extract inhibited TNF-α production in a concentration-dependent manner suggesting that it had immunosuppressive potential. Inhibition at 10 µg/mL was found to be 55% as against 48% using ß-methasone. CONCLUSION: The results suggested that BAK-I extract can be used as a potential immunosuppressive agent.

Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell (Scrophulariaceae).

BACKGROUND: Endophytes, which reside in plant tissues, have the potential to produce novel metabolites with immense benefits for health industry. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) Pennell were investigated. METHODS: Endophytic fungi were isolated from the Bacopa monnieri. Extracts from liquid cultures were tested for cytotoxicity against a number of cancer cell lines using the MTT assay. Antimicrobial activity was determined using the micro dilution method. RESULTS: 22% of the examined extracts showed potent (IC50 of <20 µg/ml) cytotoxic activity against HCT-116 cell line. 5.5%, 11%, 11% of the extracts were found to be cytotoxic for MCF-7, PC-3, and A-549 cell lines respectively. 33% extracts displayed antimicrobial activity against at least one test organism with MIC value 10-100 µg/ml. The isolate B9_Pink showed the most potent cytotoxic activity for all the cell lines examined and maximum antimicrobial activity against the four pathogens examined which was followed by B19. CONCLUSIONS: Results indicated the potential for production of bioactive agents from endophytes of Bacopa monnieri.

Field efficacy of minidosed pour-on ivermectin and eprinomectin against goat warble fly infestation by Przhevalskiana silenus.

The efficacy of minidose of pour-on ivermectin and eprinomectin formulations against first instar larvae of Przhevalskiana silenus was observed in naturally infested goats in the Jammu region, North India. The study was performed in mid August 2011. A total of 280 goats were randomly divided in to 7 groups of 40 each. Goats of the first three groups were treated with pour-on ivermectin at dosage of 2, 5, and 200 µg/kg body weight, respectively, whereas animals of the fourth to sixth groups were treated with pour-on eprinomectin at 25, 50, and 500 µg/kg body weight, respectively. Group VII animals were kept as untreated control. The results indicated that no warbles were recorded between December 2011 and March 2012 on back of animals treated with pour-on preparations of ivermectin at dosage of 5 and 200 µg/kg body weight, respectively, and eprinomectin at dosage of 50 and 500 µg/kg body weight, respectively. Thus, it is concluded that administration of minidose of pour-on ivermectin (5 µg/kg body weight) and eprinomectin (50 µg/kg body weight) is cost effective and so can be used for warble fly control campaign in Jammu region.

Biodiversity of endosymbiont fungi associated with a marine sponge Lamellodysidea herbacea and their potential as antioxidant producers.

This study aims to isolate endosymbiontic fungi from the marine sponge Lamellodysidea herbacea and to explore their antioxidant potential. Marine-derived fungi, with their vast biodiversity, are considered a promising source of novel antioxidants which can replace synthetic ones. Marine sponges have previously reported bioactive properties that could ameliorate oxidative stress, particularly their associated fungi, producing high-frequency bioactive molecules (adaptogenic molecules) in response to stressors. 19 endosymbiont fungi associated with marine sponges were isolated, and their extracts were evaluated for their antioxidant capacities. Extract of an endosymbiont fungus, isolate SPG6, identified as Alternaria destruens, through surface electron microscopy (SEM) and ITS gene sequencing, showed broad range antioxidant activities (EC50 values) (free radical scavenging 32.54 mg L-1, Hydroxyl radical scavenging activity < 0.078 g L-1, total reducing power 0.114 g L-1, Chelating power 0.262 g L-1, H2O2 scavenging activity < 0.078 g L-1, and Superoxide radical scavenging activity > 5.0 g L-1). The extract of isolate SPG6 was fractioned and analyzed through GC-MS. Marine sponge-associated endosymbiont fungi are a rich source of antioxidant molecules. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-03972-1.

Biotransformation of eugenol by an endophytic fungus Daldinia sp. IIIMF4010 isolated from Rosmarinus officinalis.

Natural value-added compounds produced from biological sources have attained immense significance in medicinal, food, flavourings, and agrochemical industries. Further, biotransformation is a powerful tool used to produce value-added compounds cost-effectively and selectively. In the present study, biotransformation of eugenol using an endophytic fungus Daldinia sp. IIIMF4010 isolated from the fresh leaves of the plant Rosmarinus officinalis leads to the production of two known value-added compounds. The biotransformation reaction of eugenol (50 mM) resulted in the production of eugenol-ß-D-glucopyranoside (6.2%) and vanillin (21.8%). These biotransformed products were further characterized by liquid chromatography-mass spectroscopy (LC-MS) and nuclear magnetic resonance (NMR).

Economic impact of Przhevalskiana silenus infestation in native goats of Northern India.

Economic losses incurred by Przhevalskiana silenus in goats of Jammu province of Jammu and Kashmir (North India) were assessed from July 2005 to June 2006. Abattoir studies revealed holes in hides for 112 days (mid-December 2005 to first week of April 2006) and trimming of carcass for 243 days (July 2005 to February 2006) during a lifecycle, leading to annual losses of Rs. 7,391,432 and Rs. 10,510,837, respectively. An effect on production traits (weight gain and milk) was assessed on naturally P. silenus-infested Beetal goats. The mean body weight loss of warbled goats was 2.40 kg (equal to 1.096 kg of meat, assuming 45.7% dressing percentage of goat) leading to an annual loss of Rs. 35,473,265. The mean milk yield loss per warbled goat per day was 101.7 g resulting in losses to the tune of Rs. 19,879,012. It can be concluded that P. silenus infestation resulted in an overall loss of Rs. 73,254,547, i.e. 73.26 million annually (equivalent to US $1.62 million; US $1 = Rs. 45.24), where reduced growth alone contributed 49% of the total losses. A study recommends that a control scheme against P. silenus infestation in goats using an effective insecticide should be followed to prevent huge economic losses in hide, milk and meat production. Economic losses associated with P. silenus infestation in goats were estimated for the first time from India.

Secondary metabolites produced by Macrophomina phaseolina, a fungal root endophyte of Brugmansia aurea, using classical and epigenetic manipulation approach.

Endophytic fungi are rich sources of structurally complex chemical scaffolds with interesting biological activities. However, their metabolome is still unknown, making them appealing for novel compound discovery. To maximize the number of secondary metabolites produced from a single microbial source, we used the "OSMAC (one strain-many compounds) approach." In potato dextrose medium, M. phaseolina produced phomeolic acid (1), ergosterol peroxide (2), and a volatile compound 1,4-benzene-diol. Incorporating an epigenetic modifier, sodium valproate, affected the metabolite profile of the fungus. It produced 3-acetyl-3-methyl dihydro-furan-2(3H)-one (3) and methyl-2-(methyl-thio)-butyrate (4), plus volatile chemicals: butylated hydroxy toluene (BHT), di-methyl-formamide, 3-amino-1-propanol, and 1,4-benzenediol, 2-amino-1-(O-methoxyphenyl) propane. The structure of compounds 1-4 was established with the help of spectroscopic data. This study revealed first-time compounds 1-4 in the fungus M. phaseolina using a classical and epigenetic manipulation approach.

Synergistic antimicrobial and antibiofilm activities of piperic acid and 4-ethylpiperic acid amides in combination with ciprofloxacin.

In the present work, piperic acid and 4-ethylpiperic acid (EPA) amides with amino acids (C1-C8) were bio-evaluated for their antimicrobial activity and biofilm inhibition against Gram-positive and Gram-negative bacterial strains. Among all, EPA-ß3,3-Pip(Bzl)-OMe, C2 displayed the potent antimicrobial activity with MIC of 6.25 µg ml-1 against Gram-negative bacteria Escherichia coli. In combination studies, the FIC indices suggested that C1 and C2 have a synergistic effect with ciprofloxacin against E. coli and Bacillus subtilis, whereas C5 exhibited a synergistic effect with ciprofloxacin against all the tested bacteria. The inhibitory effect of amides C1, C2, and C5 on the biofilm formation of test strains was significantly potentiated by co-administration with ciprofloxacin. Furthermore, the effective concentrations of C2 in combination reduced drastically compared to alone for biofilm inhibition. At these concentrations, C2 showed negligible hemolytic and cytotoxic activities.

Prevalence and biology of goat warble fly infestation by Przhevalskiana silenus in Jammu province, India.

Examination of 3,960 goats brought from six districts of Jammu province and slaughtered at Jammu abattoir from July 2005 to June 2006 revealed 47.22% prevalence of larval stages of warble fly (Przhevalskiana silenus Brauer, 1858) in goats. Highest prevalence was recorded in Udhampur (56.44%) and lowest in Doda district (13.49%). Classification of the study area into four zones based on height (meters above sea level), viz., zone I (300-325 m asl), zone II (325-800 m asl), zone III (800-1,500 m asl), and zone IV (>1,500 m asl), revealed significant (p < 0.01) difference in infestation among animals of zone II (71.68%), zone III (40.12%), and zone IV (22.41%). However, animals of zone I did not reveal any infestation. Statistical analysis in relation to age showed significant (p < 0.01) difference among different age groups, i.e., <1 year (2.81%), 1-3 years (51.17%), and >3 years (43.16%). Breed-wise analysis also showed significantly (p < 0.01) higher infestation rate among Bakerwali (51.51%) goats as compared to Beetal (42.59%). But no significant difference was recorded among male (47.81%) and female (46.82%) animals. The overall mean larva count (L1, L2, and L3) was observed to be 14.72 ± 0.34, ranging from 4 to 72. It was significantly (p < 0.05) higher in animals of zone II (22.20 ± 0.21) as compared to goats of zones III (14.21 ± 0.41) and IV (7.73 ± 0.90). Age-wise analysis of mean larva count also showed significant (P < 0.05) difference between animals of 1-3 years (16.25 ± 0.37) and >3 years of age (13.18 ± 0.40). Mean larvae count in relation to sex and breed did not reveal any significant difference. First-instar larvae (L(1)) were recorded from May to mid-September, second larval stage (L(2)) from mid-September to mid-December, and third-stage larvae (L(3)) from mid-December to February. However, No larvae were recorded from March to April, which is suggestive of pupation period in this region. Thus, it is concluded that adult fly is active in April to June. The results further confirmed that internal life cycle of P. silenus is subcutaneous and no migration of larvae occurs. This is the first report from India, based on slaughter house study on the prevalence and biology of goat warble fly infestation. It will help in devising suitable prophylactic and eradication program to check the economic losses rendered by adult fly.

Tetrahydropiperic acid (THPA) conjugated cationic hybrid dipeptides as antimicrobial agents.

The present work describes the synthesis of hybrid dipeptides H-Lys-Gpn-PEA, C1; H-Lys-ß3,3AC6C-PEA, C2, and THPA conjugated dipeptides, THPA-Lys-Gpn-PEA, C3, and THPA-Lys-ß3,3AC6C-PEA, C4. All the peptides were evaluated against both Gram-negative and Gram-positive bacterial strains. Among all, peptide C4 exhibited the most potent activity with MIC 1.56 µM against P. aeruginosa (MTCC 424) and S. aureus (MTCC 737). Further, time-kill kinetics, fluorescence assays, and scanning electron microscopy (SEM) studies were performed in order to understand the mechanism of action and efficacy of peptide C4, The fluorescence assays and SEM images demonstrated the bacterial killing through membrane disruption. The peptide C4 exhibited very low hemolytic activity with negligible cytotoxicity against normal human breast cell line FR2.