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1.  Application of immobilized tannase from Aspergillus niger for the removal of tannin from myrobalan juice 
Indian Journal of Microbiology  2010;50(Suppl 1):46-51.
Tannase produced optimally on an agroresidue by an Aspergillus niger isolate under submerged fermentation immobilized on sodium alginate beads with 93.6% efficiency was applied for tannin removal from myrobalan/aonla (Phyllanthus emblica) juice. The pH and temperature optima of the immobilized enzyme were found to be 5.4 and 40°C while the corresponding values of the soluble enzyme were 5.8 and 35°C. Maximum tannin removal of 73.6% was obtained at 40°C and 150 rpm in 180 min with 36.6 U/ml of immobilized enzyme while the same amount of the soluble enzyme removed 45.2% of tannin at 37°C and 150 rpm in the same time period. The immobilized beads could be used repeatedly till 7th cycle with 77% efficiency. When preserved at 6°C the beads retained 71.7% of enzyme activity after 60 days. Reduction in vitamin C content, which is responsible for antioxidant property of the fruit, was minimum at only 2% during the treatment.
PMCID: PMC3396397  PMID: 22815571
Aspergillus niger; Immobilized tannase; Myrobalan; Submerged fermentation; Sodium alginate
2.  Hepatoprotective effect of poly herbal formulation against various hepatotoxic agents in rats 
Pharmacognosy Research  2012;4(1):50-56.
Individually Andrographis paniculata Nees. (Acanthaceae), Phyllanthus niruri Linn.(Euphorbiaceae) and Phyllanthus emblica Linn. single plant extracts have been reported to have hepatoprotective activity. However, literature survey shows that no sufficient scientific data has been publish on pharmacological evaluation of these plants in combined form.
Hepatoprotective activity of the polyherbal hepatoprotaective formulation (PHF)-containing spray-dried aqueous extracts of Andrographis paniculata Nees. (Acanthaceae), Phyllanthus niruri Linn. (Euphorbiaceae) and Phyllanthus emblica Linn. (Euphorbiaceae), was screened against paracetamol, carbon tetrachloride (CCl4), and ethanol-induced hepatic damage in rats. PHF was evaluated by measuring levels of serum marker enzymes like SGOT, SGPT, ALP, direct bilirubin (DB), and lactate dehydrogenase (LDH). The histological studies were also studied support the biochemical parameters. Silymarin was used as standard drug.
Administration of PHF (100 and 200 mg/kg p.o.) significantly inhibited paracetamol, CCl4 and ethanol-induced elevation levels of SGPT, SGOT, ALP, DB and LDH. A comparative histopathological study of liver exhibited almost normal architecture as compared to toxicant group.
Results suggests that the hepatoprotective effects of PHF might be useful for liver protection due to combined action of all plant extracts along with their phytoconstituents.
PMCID: PMC3250040  PMID: 22224062
Andrographis paniculata Nees; carbon tetrachloride; hepatoprotective activity; Marker enzymes; paracetamol; Phyllanthus niruri Linn
3.  Quantification of Gallic Acidin Fruits of Three Medicinal Plants 
Triphala is a traditional herbal formulation consisting of dried fruits originating from three medicinal plants, namely Terminalia chebula, Terminalia bellerica and Phyllanthus emblica. It is used in folk medicine for the treatment of headaches, dyspepsia and leucorrhoea. There are some reports regarding Triphala’s pharmacological effects including its anti-cancer, radioprotective, hypocholesterolaemic, hepatoprotective and anti-oxidant activities. The most important components of these plants are the tannins and gallic acid which they contain. Gallic acid being a compound with tannin structure existing in the Triphala fruit. In this research, the gallic acid content contained in the three plants constituting Triphala was determined. Plant fruits were purchased from available Iranian markets. Milled and powdered fruits from each plant were extracted with 70% acetone and subjected to a reaction with rhodanine reagent in the process forming a colored complex. The complex’s absorbance was measured at 520 nm and the amount of gallic acid was determined using its calibration curve. According to the results, the highest amount of gallic acid was observed in Phyllanthus embelica (1.79-2.18%) and the lowest amount was found in Terminalia chebula (0.28-0.80%). Moreover, differences between plant samples from different markets places were found to be statistically significant (p < 0.05). These differences can possibly be due to the source of plant preparation, storage condition and period of Triphala storage. In general, the rhodanine assay is a simple, rapid and reproducible method for the standardization of Triphala as gallic acid.
PMCID: PMC3828909  PMID: 24250348
Triphala; Terminalia chebula; Terminalia bellerica; Phyllanthus emblica; Gallic acid; Rhodanine assay
4.  RAPD based genetic variability among cultivated varieties of Aonla (Indian Gooseberry, Phyllanthus emblica L.) 
Aonla, the Indian Gooseberry (Phyllanthus emblica) is widely grown in India due to its neutraceutical properties. Investigations on the use of RAPD markers enabled us to estimate genetic variability among commercially cultivated varieties. This study also enabled us to distinguish these varieties using a set of four decamer primers, which was otherwise difficult by using morphological markers. Cluster analysis revealed three different groups of varieties directly associated to their place of origin. RAPD markers were also able to differentiate varieties of same origin or even selection from same parents. This information can be used for identification of varieties and further crop improvement programme.
PMCID: PMC3550374  PMID: 23572926
Aonla; Phyllanthus emblica; RAPD
5.  Antioxidant effect ofPhyllanthus emblica fruits on healing of indomethacin induced gastric ulcer in rats 
Post-treatment of the indomethacin induced ulcerated rats at the optimal dose of 100 mg/kg body-wt. orally for 7 consecutive days with the lyophilized aqueous extract of the fruits ofPhyllanthus emblica L. syn.Emblica officinalis Gaertn. (Euphorbiaceae) exhibited highly significant (p<0.001) enhancement of secretion of catalase, reduced glutathione and decrease in malonyldialdehyde (MDA). Furthermore, the gross morphological observation and highly significant (p<0.001) decrease of ulcer index (81.43%) indicated healing effect of the extract on gastric ulcer.
PMCID: PMC3453744  PMID: 23105366
Antioxidant; antiulcer; healing; Phyllanthus emblica
6.  Effects of Phyllanthus emblica extract on endothelial dysfunction and biomarkers of oxidative stress in patients with type 2 diabetes mellitus: a randomized, double-blind, controlled study 
It has been reported that hyperglycemia can induce endothelial dysfunction via increased oxidative stress and that it plays a central role in the development of atherosclerosis and coronary heart disease. Phyllanthus emblica (Emblica officinalis, amla) is known for its antioxidant and antihyperlipidemic activity. The present study compared the effects of an aqueous extract of P. emblica (highly standardized by high-performance liquid chromatography to contain low molecular weight hydrolyzable tannins, ie, emblicanin A, emblicanin B, pedunculagin, and punigluconin) versus those of atorvastatin and placebo on endothelial dysfunction and biomarkers of oxidative stress in patients with type 2 diabetes.
Eligible patients were randomized to receive either P. emblica 250 mg twice daily, P. emblica 500 mg twice daily, atorvastatin 10 mg in the evening and matching placebo in the morning, or placebo twice daily for 12 weeks. The primary efficacy parameter was the change in endothelial function identified on salbutamol challenge at baseline and after 12 weeks of treatment. Secondary efficacy parameters were changes in biomarkers of oxidative stress (malondialdehyde, nitric oxide, and glutathione), high sensitivity C-reactive protein levels, the lipid profile, and glycosylated hemoglobin (HbA1c) levels. Laboratory safety parameters were measured at baseline and after 12 weeks of treatment.
Eighty patients completed the study. Treatment with P. emblica 250 mg, P. emblica 500 mg, or atorvastatin 10 mg produced significant reductions in the reflection index (−2.25%±1.37% to −9.13%±2.56% versus −2.11%±0.98% to −10.04%±0.92% versus −2.68%±1.13% to −11.03%±3.93%, respectively), suggesting improvement in endothelial function after 12 weeks of treatment compared with baseline. There was a significant improvement in biomarkers of oxidative stress and systemic inflammation compared with baseline and placebo. Further, the treatments significantly improved the lipid profile and HbA1c levels compared with baseline and placebo. All treatments were well tolerated.
Both atorvastatin and P. emblica significantly improved endothelial function and reduced biomarkers of oxidative stress and systemic inflammation in patients with type 2 diabetes mellitus, without any significant changes in laboratory safety parameters.
PMCID: PMC3735284  PMID: 23935377
Phyllanthus emblica; atorvastatin; endothelial dysfunction; type 2 diabetes
7.  Effect of zero energy cool chamber and post-harvest treatments on shelf-life of fruits under semi-arid environment of Western India. Part 2. Indian gooseberry fruits 
Effect of zero energy cool chamber (ZECC) along with post-harvest treatments including CaCl2, mustard oil and K2SO4 separately on shelf-life and fruit quality attributes of Indian gooseberry or aonla (Emblica officinalis Gaertn) during storage under semi-arid ecosystem of Gujarat was studied. Increase in physiological loss in weight (PLW), spoilage loss, total soluble solids, total sugar and reducing sugars, reduction in titratable acidity and ascorbic acid were observed during storage period in all the treatments. Fruits treated with 1.5% CaCl2 and stored in ZECC recorded least PLW (16%), spoilage loss (16.5%), respiratory activity (83 mg CO2 /kg/h) and exhibited 11 days of shelf-life, while untreated control had 6 days economic life. It was closely followed by 1% CaCl2 + ZECC treatment. Fruits stored in ZECC recorded 9 days shelf-life. Highest respiration rate was in control (88.1 mg CO2 /kg/h) on 13th day of storage. It may be concluded that 1.5% CaCl2 and storage in ZECC treatment was found most efficient to retain the fruit quality attributes till 13th day of storage under semi-arid environment of western India.
PMCID: PMC3551009  PMID: 23572669
Aonla; Emblica officinalis; Zero energy cool chamber; Calcium chloride; Shelf-life; Spoilage
8.  Development and quality evaluation of aonla mouth freshner 
Nutritive and palatable mouth freshners were prepared from dehydrated aonla (Emblica officinalis Gaertn) pulp of ‘Desi’ and ‘Banarsi’ cultivars by mixing carboxy methyl cellulose, gums, arecanut, cardamom, sugar and milk powder at different proportions as a substitute for pan masala, tobacco and gutka. Mouth fresheners developed were packed in high density polyethylene pouches (HDPE, 100 gauge), stored at ambient conditions (8–20 °C, 60%RH) and analysed for physico-chemical and sensory quality attributes at different storage intervals. During storage for 6 months, ascorbic acid and overall acceptability of mouth freshener decreased (p ≤ 0.05) and moisture content increased. The equivalent relative humidity of mouth freshener was 49% and 53% in ‘Desi’ and ‘Banarsi’ cultivars, respectively. Despite the changes observed in various physico- chemical and sensory attributes, the overall sensory quality attributes of mouth freshners remained acceptable.
PMCID: PMC3551145  PMID: 23572710
Aonla; Mouth freshener; Ascorbic acid; Sensory acceptability
9.  Determination of gallic acid in Phyllanthus emblica Linn. dried fruit powder by HPTLC 
Emblica (Phyllanthus emblica L.), an euphorbiaceous plant, is widely distributed in subtropical and tropical areas of India, China and Indonesia. The fruits possess antimicrobial, antioxidant, anti-inflammatory, analgesic and antipyretic properties. In the current article a new, simple, sensitive, selective, precise, and robust high-performance thin-layer chromatographic (HPTLC) method was developed and validated for the determination of gallic acid in dried fruit powder of Phyllanthus emblica.
Materials and Methods:
The quantitative determination of gallic acid was performed on TLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase. The linear ascending development was carried out in a twin trough glass chamber saturated with a mobile phase consisting of toluene: ethyl acetate: formic acid: methanol (3:3:0.8:0.2) at room temperature (25 ± 2°C). Camag TLC scanner III was used for spectrodensitometric scanning and analysis, in the absorbance mode, at 278 nm.
The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.99977 in the concentration range of 40 – 240 ng spot—1, with respect to the peak area. According to the guidelines of the International Conference on Harmonization (ICH), the method was validated for precision, accuracy, and recovery.
Statistical analysis of the data showed that the method was reproducible and selective for the estimation of gallic acid.
PMCID: PMC3147091  PMID: 21814441
Phyllanthus emblica; HPTLC fingerprinting; gallic acid
10.  Fungi and Mycotoxins in Feed Intended for Sows at Different Reproductive Stages in Argentina 
The aim of this study was to evaluate fungi and contamination levels of aflatoxin B1, ochratoxin A, fumonisin B1, and zearalenone in raw materials and finished feed intended for sows at different reproductive stages. Total fungi, Aspergillus, Penicillium, and Fusarium species occurrence, were examined. Aspergillus flavus, A. niger aggregate spp., and F. verticillioides were the prevalent species. Fungal counts exceeded the levels proposed as feed hygienic quality limits (1 × 104 colony forming units) at all reproductive stages. Aflatoxin B1, ochratoxin A, fumonisin B1, and zearalenone were detected by high-pressure liquid chromatography. Aflatoxin levels in 80% samples of finished sow feeds were over the permitted levels of 0.02 μg g−1 (mean 228.2 ± 95 μg Kg−1). Fumonisin B1 was detected in all tested raw materials at levels that varied from 50.3 to 1137.64 μg Kg−1 and finished feed samples at levels that ranged from 99.8 to 512.4 μg Kg−1. Aflatoxin B1, zearalenone, and ochratoxin A were not detected in raw materials. All finished feeds were negative for zearalenone contamination whereas all nonpregnant gilt samples were contaminated with low OTA levels (mean 0.259 ± 0.123). This fact requires periodic monitoring to prevent the occurrence of mycotoxicosis in animal production, to reduce the economic losses, and to minimize hazards to human health.
PMCID: PMC2896851  PMID: 20613957
11.  Clinical study of the Immunoglobululin Enhancing effect of “Bala compound” on Infants 
Ancient Science of Life  2009;28(3):18-22.
Kaumarbhritya a branch of Asthanga Ayurveda deals with neonatal, infant and child health care. Multicentric studies conducted in various developed and developing countries have indicated that Infant Mortality Rate (I.M.R.) is very high in developing countries, and infection has been observed as the major cause. Immune system in neonates is not yet fully functional. Bala compound having the ingredients of Atibala (Abutilon indicum Linn), Amalaki (Emblica officinalis Linn), Vidanga (Emblica ribes burn), Guduchi (Tinospora cordifolia Welld Miers), Pippali (Piperlongum linn), Yashtimadhu (Glycyrrhiza glabra Linn), Shankhapuspi (Convolvulus pluricaulis Chois ), Vacha (Acorus calamus Linn), Musta (Cyperus rotundus Linn) and Ativisha (Aconitum heterophyllum wall) are Medhya as well as Rasayana drugs mention in Ayurvedic classics. ‘Bala compound” was tried in infants in the form of oral drops for a period of six months and result was assessed for serum immuoglobulins IgG, IgM, IgA for three months of interval of two follow ups (i.e., third and six month of infant). There is significant increase of immunoglobulins observed after six months administration of ‘Bala compoumd”
PMCID: PMC3336317  PMID: 22557316
12.  Loss of msnA, a Putative Stress Regulatory Gene, in Aspergillus parasiticus and Aspergillus flavus Increased Production of Conidia, Aflatoxins and Kojic Acid 
Toxins  2011;3(1):82-104.
Production of the harmful carcinogenic aflatoxins by Aspergillus parasiticus and Aspergillus flavus has been postulated to be a mechanism to relieve oxidative stress. The msnA gene of A. parasiticus and A. flavus is the ortholog of Saccharomyces cerevisiae MSN2 that is associated with multi-stress response. Compared to wild type strains, the msnA deletion (∆msnA) strains of A. parasiticus and A. flavus exhibited retarded colony growth with increased conidiation. The ∆msnA strains also produced slightly higher amounts of aflatoxins and elevated amounts of kojic acid on mixed cereal medium. Microarray assays showed that expression of genes encoding oxidative stress defense enzymes, i.e., superoxide dismutase, catalase, and cytochrome c peroxidase in A. parasiticus ∆msnA, and the catalase A gene in A. flavus ∆msnA, was up-regulated. Both A. parasiticus and A. flavus ∆msnA strains produced higher levels of reactive oxygen species (ROS), and ROS production of A. flavus msnA addback strains was decreased to levels comparable to that of the wild type A. flavus. The msnA gene appears to be required for the maintenance of the normal oxidative state. The impairment of msnA resulted in the aforementioned changes, which might be used to combat the increased oxidative stress in the cells.
PMCID: PMC3210457  PMID: 22069691
Aspergillus; aflatoxin; kojic acid; oxidative stress; development
13.  RAPD Analysis for Determination of Components in Herbal Medicine 
In this study, the RAPD (Random Amplified Polymorphic DNA) technique was employed for determination of the components in an Ayurvedic herbal prescription, Rasayana Churna. One-hundred-and-twenty decamer oligonucleotide primers were screened in the RAPD analysis to identify three Ayurvedic medicines, dried stem of Tinospora cordifolia, dried fruit of Emblica officinalis and dried fruit of Tribulus terestris, the Ayurvedic prescription. Primer OPC-6 simultaneously generated three distinct amplicons, each specific to one component. The marker with 600 bp is specific to Tinospora cordifolia; the marker 500 bp is specific to Emblica officinalis and the remaining marker >1000 bp was present in Tribulus terestris. Presence of three herbal medicines was determined when RAPD reaction with OPC-6 was performed. The technique was proved to contribute to the identification of components in Ayurvedic herbal preparation and thus helping to serve as a complementary tool for quality control.
PMCID: PMC2206231  PMID: 18227927
DNA fingerprinting; herbal medicine; RAPD; standardization
14.  Implications of fungal infections and mycotoxins in camel diseases in Saudi Arabia 
Natural feed ingredients (corn, barley and wheat bran) and compound feed (manufactured pellet) are two types of fodder used for animal feeding, especially camel in Saudi Arabia. Twenty samples of each type of fodder were collected from seven different regions and screened for the presence of fungi, aflatoxins, ochratoxin and zearalenone. Fungal isolation of natural feed ingredients yielded 10 genera and 38 species of different fungi. Compound fodder samples were contaminated with 16 genera and 32 species of fungi. Total counts of Aspergillus, Penicillium and Fusarium in the animal feed samples were ranged from 54 to 223 × 103, 31.9 to 60 × 103 and 18 to 29 × 103 CFU/g, respectively. These isolates when tested for aflatoxin, ochratoxin and zearalenone producing ability, revealed this property in only four isolate, identified as Aspergillus flavus, A. parasiticus, A. ochraceus and Fusarium graminaerum. The percentage of toxigenic fungi was ranged from 5.5% to 30% for natural feed ingredients and from 4.5% to 20% for compound feed. The incidence of aflatoxins (AFT) in samples of natural feed ingredients was found to be ranged from 1 to 24.8 ppb, ochratoxin A (OTA) ranged from 1 to 44 ppb and zearalenone (ZON) ranged from 1 to 23 ppb. Contamination of compound feed with aflatoxin and ochratoxin A was ranged from 1 to 6.4 ppb and 1 to 4.7 ppb, respectively. All samples collected were found contaminated with fungi or their toxins and natural feed samples were more contaminated compared to compound feed samples. The concentrations detected were in the allowed limit (<20 ppb) except four samples of natural feed ingredients which were above the allowed limit of the tested mycotoxins. In conclusion, feed samples were contaminated with fungi and some toxigenic isolates which were responsible about mycotoxin production. Some samples had exceeded amount of AFT, OTA and ZON and may be contaminated with other mycotoxins which mean implication of fungi in camel health problems and death in Saudi Arabia.
PMCID: PMC3730929  PMID: 23961061
Mycotoxin; Aflatoxins; Ochratoxin A; Zearalenone; Aspergillus; Camel feed; Fodder; Fungal infection
15.  Phyllanthus emblica L. Enhances Human Umbilical Vein Endothelial Wound Healing and Sprouting 
Endothelial dysfunction is the hallmark of impaired wound healing and increased risk of cardiovascular disease. Antioxidants from natural sources decrease oxidative stress and protect against cellular damage caused by reactive oxygen species (ROS). In this study, we examined the antioxidant constituents and capacity of Phyllanthus emblica L. (PE) fruit in freeze-dried power form. The pharmacological properties of PE were investigated using human umbilical vein endothelial cells (HUVECs) in the aspects of endothelial cell proliferation, nitric oxide (NO) production, wound healing, cell migration, in vitro angiogenesis, and VEGF gene expression. The ASC content of PE was 1.574% + 0.046% (w/w) as determined by HPLC and the total phenolic content was 36.1% ± 0.7% gallic acid equivalent when measured by Folin-Ciocalteu assay. The FRAP assay revealed a relatively high antioxidant capacity at 3,643 + 192.5 µmole/mg. PE at 0.1 to 10 µg/mL did not significantly influence endothelial cell proliferation, but at higher concentrations PE decreased cell survival to 62%. PE significantly promoted NO production, endothelial wound closure, endothelial sprouting, and VEGF mRNA expression. Therefore, PE is a candidate for antioxidant supplement that promotes endothelial function and restores wound healing competency.
PMCID: PMC3626238  PMID: 23606890
16.  Evaluation of STAT5A Gene Expression in Aflatoxin B1 Treated Bovine Mammary Epithelial Cells 
Advanced Pharmaceutical Bulletin  2013;3(2):461-464.
Purpose: Aflatoxin B1 (AFB1) is a potent mycotoxin which has been produced by fungi such as Aspergillus flavus and Aspergillus parasiticus as secondary metabolites due to their growth on food stuffs and induces hepatocellular carcinoma in many animal species, including humans. In the present study, the effect of AFB1 on STAT5A gene expression was investigated in bovine mammary epithelial cells using real time RT-PCR. Methods: Bovine mammary epithelial cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, cells were treated with AFB1 and incubated for 8 h. For real time PCR reaction, total RNA from the cultured and treated cells was extracted and used for complementary DNA synthesis. Results: The expression of STAT5A gene was significantly down regulated by AFB1 in dose- dependent manner and led to the reduction of proliferation and differentiation of epithelial cells, which has direct effect in milk protein quantity and quality. Conclusion: According to the results, it seems that down regulation of STAT5A gene in AFB1-treated cells maybe due to DNA damage induced by AFB1 in bovine mammary epithelial cells.
PMCID: PMC3848230  PMID: 24312879
Matrigel; Aflatoxin B1; 3D cell culture; STAT5A; Epithelial cells
17.  Antimicrobial Effects of Ionizing Radiation on Artificially and Naturally Contaminated Cacao Beans 
With an initial microbial level of ca. 107 microorganisms per g of Ivory Coast cacao beans, 5 kGy of gamma radiation under an atmosphere of air reduced the microflora per g by 2.49 and 3.03 logs at temperatures of 35 and 50°C, respectively. Bahia cacao beans were artificially contaminated with dried spores of Aspergillus flavus and Penicillium citrinum, giving initial fungal levels of 1.9 × 104 and 1.4 × 103 spores per g of whole Bahia cacao beans, respectively. The average D10 values for A. flavus and P. citrinum spores on Bahia cacao beans were 0.66 and 0.88 kGy, respectively.
PMCID: PMC239785  PMID: 16346530
Ancient Science of Life  2007;26(3):40-44.
Pharmacognostical and preliminary phytochemical studies of Triphala churnam were carried out. The churnam of triphala consists of equal quantities of deseeded fruits of Terminalia chebula, Terminalia bellerica and Emblica officinalis. Triphala is exclusively used in more than 200 drug formulations in Indian system of Medicine. The present study involved the pharmacognostical evaluation of Triphala, in which morphological and powder microscopical characters were established. In addition, physico-chemical parameters such as ash values viz, total ash (10.21± 0.42), acid insoluble ash (2.54 ± 0.06), water-soluble ash (5.46±0.24) and sulphated ash (13.12 ± 0.63), extractive values viz, alcohol soluble extractive (11.20±0.18)) and water-soluble extractive (52.56±2.04), fluorescent analysis and microchmical tests were determined. The preliminary phytochemical study revealed the presence of carbohydrates, reducing sugar and tannins in aqueous extract and carbohydrates, flavonoids and tannins in alcoholic extract. This standardization would be very much helpful for the identification of Triphala churnam to differentiate from other powdered sources.
PMCID: PMC3330880  PMID: 22557240
19.  Direct visual detection of aflatoxin synthesis by minicolonies of Aspergillus species. 
Single-spore colonies of Aspergillus flavus and Aspergillus parasiticus, grown for 4 to 5 days at 25 degrees C on a coconut extract agar containing sodium desoxycholate as a growth inhibitor, produced aflatoxin, readily detectable as blue fluorescent zones under long-wave (365 nm) UV light. Over 100 colonies per standard petri dish were scored for aflatoxin production by this procedure. Progeny from some strains remained consistently stable for toxin production after repeated subculture, whereas instability for toxin synthesis was revealed among progeny from other strains. Spore color markers were used to rule out cross-contamination in monitoring strains. A yellow-spored and nontoxigenic strain of A. flavus, reported previously to produce aflatoxin in response to cycloheximide treatment, proved to be toxin negative even after repeated exposure to cycloheximide. Extended series of progeny from another strain of A. flavus and from a strain of A. parasiticus were each compared by this plating procedure and by fluorometric analysis for aflatoxin when grown in a coconut extract broth. Both of these strains showed variation for toxin synthesis among their respective progeny, and specific progeny showed a good correlation for aflatoxin synthesis when examined by the two procedures.
PMCID: PMC202954  PMID: 2504116
20.  Mycotoxin-Producing Ability and Chemotype Diversity of Aspergillus Section Flavi from Soils of Peanut-Growing Regions in Iran 
Indian Journal of Microbiology  2012;52(4):551-556.
Invasion of crops with Aspergillus flavus may result in contamination of food and feed with carcinogenic mycotoxins such as aflatoxins (AF) and cyclopiazonic acid (CPA). In the present study, distribution and toxigenicity of Aspergillus flavus and A. parasiticus in soils of five peanut fields located in Guilan province, Northern Iran was investigated. From a total of 30 soil samples, 53 strains were isolated which all of them were finally identified as A. flavus by a combination of colony morphology, microscopic criteria and mycotoxin profiles. Chromatographic analysis of fungal cultures on yeast extract sucrose broth by tip culture method showed that 45 of the 53 A. flavus isolates (84.9 %) were able to produce either CPA or AFB1, while eight of the isolates (15.1 %) were non-toxigenic. The amounts of CPA and AFB1 produced by the isolates were reported in the range of 18.2–403.8 μg/g and 53.3–7446.3 μg/g fungal dry weights, respectively. Chemotype classification of A. flavus isolates based on the ability for producing mycotoxins and sclerotia showed that 43.4 % were producers of CPA, AFB1 and sclerotia (group I), 13.2 % of CPA and AFB1 (group II), 9.4 % of AFB1 and sclerotia (group III), 13.2 % of AFB1 (group IV), 5.7 % of CPA and sclerotia (group V) and 15.1 % were non-toxigenic with no sclerotia (group VI). No strain was found as producer of only CPA or sclerotia. These results indicate different populations of mycotoxigenic A. flavus strains enable to produce hazardous amounts of AFB1 and CPA are present in peanuts field soils which can be quite important regard to their potential to contaminate peanuts as a main crop consumed in human and animal nutrition.
PMCID: PMC3516660  PMID: 24293709
Aspergillus flavus; Cyclopiazonic acid; Aflatoxin; Chemotype diversity; Soil; Peanuts
21.  Mycotoxin-Producing Potential of Mold Flora of Dried Beans 
Applied Microbiology  1975;29(4):522-526.
To evaluate the potential for mycotoxin production by molds in dried beans, the mold flora of 114 samples was determined both before and after surface disinfection of the beans with 5% NaOCl. Surface disinfection substantially reduced mold incidence, indicating that contamination was mainly on the surface. The flora, both before and after disinfection, was dominated by species of the Aspergillus glaucus group, the toxicogenic species A. ochraceus, Penicillium cyclopium, and P. viridicatum, and species of Alternaria, Cladosporium, and Fusarium. The toxicogenic species Aspergillus flavis, A. versicolor, Penicillium citrinum, P. expansum, P. islandicum, and P. urticae were encountered less frequently. Of 209 species of Aspergillus and Penicillium screened for mycotoxin production on sterile rice substrate, 114 produced one or more of the following mycotoxins: A. flavus, aflatoxins; A. ochraceus, ochratoxins; A. nidulans, A. unguis, and A. versicolor, sterigmatocystin; P. cyclopium, penicillic acid; P. citrinum and P. viridicatum, citrinin; P. urticae, patulin and griseofulvin. Sterigmatocystin production by A. unguis is reported for the first time.
PMCID: PMC187018  PMID: 1168442
22.  Fungi and Mycotoxins from Pre- and Poststorage Brewer's Grain Intended for Bovine Intensive Rearing 
ISRN Veterinary Science  2012;2012:396590.
The aim of the study was to determine the mycobiota and natural levels of mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA), fumonisin B1 (FB1), and deoxynivalenol (DON) present in brewers grains pre- and poststored intended for bovine intensive rearing. Poststored (80%) samples had counts higher than 1 × 104 colony-forming units (CFU/g). Cladosporium spp. and Aspergillus spp. were isolated at high frequencies. Aspergillus flavus was the prevalent isolated species. Prestored (70%) and poststored (100%) samples showed AFB1 levels over the recommended limits (20 μg/Kg), and OTA levels were below the recommended limits (50 μg/Kg) while pre- and poststored samples did not show FB1 and DON natural contamination levels. The presence of mycotoxins in this substrate indicates the existence of contamination. Regular monitoring of feeds is required in order to prevent chronic and acute toxic syndromes related to this kind of contamination.
PMCID: PMC3671735  PMID: 23762582
23.  Inhibition of Aflatoxin Production by Surfactants 
The effect of 12 surfactants on aflatoxin production, growth, and conidial germination by the fungus Aspergillus flavus is reported. Five nonionic surfactants, Triton X-100, Tergitol NP-7, Tergitol NP-10, polyoxyethylene (POE) 10 lauryl ether, and Latron AG-98, reduced aflatoxin production by 96 to 99% at 1% (wt/vol). Colony growth was restricted by the five nonionic surfactants at this concentration. Aflatoxin production was inhibited 31 to 53% by lower concentrations of Triton X-100 (0.001 to 0.0001%) at which colony growth was not affected. Triton X-301, a POE-derived anionic surfactant, had an effect on colony growth and aflatoxin production similar to that of the five POE-derived nonionic surfactants. Sodium dodecyl sulfate (SDS), an anionic surfactant, and dodecyltrimethylammonium bromide, a cationic surfactant, suppressed conidial germination at 1% (wt/vol). SDS had no effect on aflatoxin production or colony growth at 0.001%. The degree of aflatoxin inhibition by a surfactant appears to be a function of the length of the hydrophobic and hydrophilic chains of POE-derived surfactants.
PMCID: PMC201276  PMID: 16349144
24.  Aflatoxins, hepatocellular carcinoma and public health 
Hepatocellular carcinoma (HCC) is one of the leading causes of cancer deaths worldwide, primarily affecting populations in the developing countries. Aflatoxin, a food contaminant produced by the fungi Aspergillus flavus and Aspergillus parasiticus, is a known human carcinogen that has been shown to be a causative agent in the pathogenesis of HCC. Aflatoxin can affect a wide range of food commodities including corns, oilseeds, spices, and tree nuts as well as milk, meat, and dried fruit. Many factors affect the growth of Aspergillus fungi and the level of aflatoxin contamination in food. Drought stress is one of the factors that increase susceptibility of plants to Aspergillus and thus aflatoxin contamination. A recent drought is thought to be responsible for finding of trace amounts of aflatoxin in some of the corn harvested in the United States. Although it’s too soon to know whether aflatoxin will be a significant problem, since United States is the world’s largest corn producer and exporter, this has raised alarm bells. Strict regulations and testing of finished foods and feeds in the United States should prevent a major health scare, and prevent human exposure to deleterious levels of aflatoxin. Unfortunately, such regulations and testing are not in place in many countries. The purpose of this editorial is to summarize the current knowledge on association of aflatoxin and HCC, encourage future research and draw attention to this global public health issue.
PMCID: PMC3602466  PMID: 23539499
Aflatoxins; Hepatocellular carcinoma; Environmental health; Food safety; Public health
25.  Effect of pretreatment and drying methods on quality of value-added dried aonla (Emblica officinalis Gaertn) shreds 
Value added dried Indian gooseberry (aonla) shreds were prepared using aonla fruits of cv. ‘NA-7’. Two blanching methods (hot water and potassium metabisulphite (KMS) at 0.1%) and two drying methods (solar and hot air oven drying) were tried for the production of aonla shreds. Common salt, black salt and ginger juice were mixed for enhancing sensory quality of the product. The best product was obtained with KMS blanching and drying in solar dryer with added common salt at 3%. The most acceptable product had ascorbic acid content 298.3 mg/100 g, tannin 2.4%, acidity 2.6%, reducing sugar 3.0%, non-reducing sugar 21.0% and total sugar 24.0%. The recovery was 8.0–8.5%.
PMCID: PMC3551091  PMID: 23572715
Aonla; Blanching; Drying; Recipe; Quality; Recovery

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