Effects of Fermentation Techniques on the Quality of Cocoa Beans

Fermentation as an important operation in the processing of cocoa beans is now affected by the recent climate change across the globe. The major requirement for effective fermentation is the ability of the material used to retain sufficient heat for the required microbial activities. Apart from the effects of climate on the rate of heat retention, the materials used for fermentation plays an important role. Most Farmers still restrict fermentation activities to the use of traditional methods. Improving on cocoa fermentation in this era of climate change makes it necessary to work on other materials that can be suitable for cocoa fermentation. Therefore, the objective of this study was to determine the effects of fermentation techniques on the quality of cocoa beans. The materials used in this fermentation research were heap-leaves (traditional), stainless steel, plastic tin, plastic basket and wooden box. The period of fermentation varies from zero days to 10 days. Physical and chemical tests were carried out for variables in quality determination in the samples. The weight per bean varied from 1.0-1.2 g after drying across the samples and the major color of the dry beans observed was brown except with the samples from stainless steel. The moisture content varied from 5.5-7%. The mineral content and the heavy metals decreased with increase in the fermentation period. A wooden box can conclusively be used as an alternative to heap-leaves as there was no significant difference in the physical features of the samples fermented with the two methods. The use of a wooden box as an alternative for cocoa fermentation is therefore recommended for cocoa farmers.

The Evaluation of Antioxidant and Antimicrobial Activities of Essential Oil and Aqueous, Methanol, Ethanol, Ethyl Acetate and Acetone Extract of Hypericum scabrum

Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl3) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Impact of Long Term Application of Municipal Solid Waste on Physicochemical and Microbial Parameters and Heavy Metal Distribution in Soils in Accordance to Its Agricultural Uses

Municipal Solid Waste (MSW), being a rich source of organic materials, can be used for agricultural applications as an important source of nutrients for soil and plants. This is also an alternative beneficial management practice for MSW generated in developing countries. In the present study, MSW treated soil samples from last four to six years at farmer’s field in Rohtak and Gurgaon states (Haryana, India) were collected. The samples were analyzed for all-important agricultural parameters and compared with the control untreated soil samples. The treated soil at farmer’s field showed increase in total N by 48 to 68%, P by 45.7 to 51.3%, and K by 60 to 67% compared to untreated soil samples. Application of sewage sludge at different sites led to increase in microbial biomass C by 60 to 68% compared to untreated soil. There was significant increase in total Cu, Cr, Ni, Fe, Pb, and Zn in all sewage sludge amended soil samples; however, concentration of all the metals were still below the current permitted (EU) limits. To study the adverse effect of heavy metals accumulation on various soil microbial activities, the sewage sludge samples (from wastewater treatment plant at Gurgaon) were artificially contaminated with heavy metal concentration above the EU limits. They were then applied to soil samples with different rates (0.5 to 4.0%) and incubated for 90 days under laboratory conditions. The samples were drawn at different intervals and analyzed for various parameters like pH, EC, total N, P, K, microbial biomass C, carbon mineralization, and diethylenetriaminepentaacetic acid (DTPA) exactable heavy metals. The results were compared to the uncontaminated sewage sludge. The increasing level of sewage sludge from 0.5 to 4% led to build of organic C and total N, P and K content at the early stages of incubation. But, organic C was decreased after 90 days because of decomposition of organic matter. Biomass production was significantly increased in both contaminated and uncontaminated sewage soil samples, but also led to slight increases in metal accumulation and their bioavailability in soil. The maximum metal concentrations were found in treatment with 4% of contaminated sewage sludge amendment.

Phenolic Compounds and Antimicrobial Properties of Pomegranate (Punica granatum) Peel Extracts

In recent years, tendency to use of natural antimicrobial agents in food industry has increased. Pomegranate peels containing phenolic compounds and anti-microbial agents, are counted as valuable source for extraction of these compounds. In this study, the extraction of pomegranate peel extract was carried out at different ethanol/water ratios (40:60, 60:40, and 80:20), temperatures (25, 40, and 55 ˚C), and time durations (20, 24, and 28 h). The extraction yield, phenolic compounds, flavonoids, and anthocyanins were measured. ‎Antimicrobial activity of pomegranate peel extracts were determined against some food-borne ‎microorganisms such as Salmonella enteritidis, Escherichia coli, Listeria monocytogenes, ‎‎Staphylococcus aureus, Aspergillus niger, and Saccharomyces cerevisiae by agar diffusion and MIC methods. Results showed that at ethanol/water ratio 60:40, 25 ˚C and 24 h maximum amount of phenolic compounds ‎‏(‎‏‎349.518‎‏ ‏mg gallic acid‏/‏g dried extract), ‎flavonoids (250.124 mg rutin‏/‏g dried extract), anthocyanins (252.047 ‎‏‏mg ‎cyanidin‏‎3‎‏glucoside‏/‏‎100 g dried extract), and the strongest antimicrobial activity were obtained. ‎All extracts’ antimicrobial activities were demonstrated against every tested ‎‎microorganisms‏.‎‏ Staphylococcus aureus showed the highest sensitivity among the tested ‎‎‎microorganisms.

The Potential Effect of Biochar Application on Microbial Activities and Availability of Mineral Nitrogen in Arable Soil Stressed by Drought

Application of biochar to arable soils represents a new approach to restore soil health and quality. Many studies reported the positive effect of biochar application on soil fertility and development of soil microbial community. Moreover biochar may affect the soil water retention, but this effect has not been sufficiently described yet. Therefore this study deals with the influence of biochar application on: microbial activities in soil, availability of mineral nitrogen in soil for microorganisms, mineral nitrogen retention and plant production. To demonstrate the effect of biochar addition on the above parameters, the pot experiment was realized. As a model crop, Lactuca sativa L. was used and cultivated from December 10th 2014 till March 22th 2015 in climate chamber in thoroughly homogenized arable soil with and without addition of biochar. Five variants of experiment (V1 – V5) with different regime of irrigation were prepared. Variants V1 – V2 were fertilized by mineral nitrogen, V3 – V4 by biochar and V5 was a control. The significant differences were found only in plant production and mineral nitrogen retention. The highest content of mineral nitrogen in soil was detected in V1 and V2, about 250 % in comparison with the other variants. The positive effect of biochar application on soil fertility, mineral nitrogen availability was not found. On the other hand results of plant production indicate the possible positive effect of biochar application on soil water retention.

Antimicrobial, Antioxidant and Free Radical Scavenging Activities of Essential Oils Extracted from Six Eucalyptus Species

Eucalyptus species are well reputed for their traditional use in Asia as well as in other parts of the world; therefore, the present study was designed to investigate the antimicrobial and antioxidant activities associated with essential oils from different Eucalyptus species. Essential oils from the leaves of six Eucalyptus species, including: Eucalyptus woodwardi, Eucalyptus stricklandii, Eucalyptus salubris, Eucalyptus sargentii, Eucalyptus torquata and Eucalyptus wandoo were separated by hydrodistillation and dried over anhydrous sodium sulphate. DPPH, ferric reducing antioxidant power, and hydroxyl radical scavenging activity assays were carried out to evaluate the antioxidant potential of the oils. The results indicate that examined oils exhibit substantial antioxidant activities relative to ascorbic acid. Previously, these oils were evaluated for their antimicrobial activities, against wide range of bacterial and fungal strains, and they were shown to possess significant antimicrobial activities. In this study, further investigation into the growth kinetics of oil-treated microbial cultures was conducted. The results clearly demonstrate that the microbial growth was markedly inhibited when treated with sub-MIC concentrations of the oils. Taken together, the results obtained indicate a high potential of the examined essential oils as bioactive oils, for nutraceutical and medical applications, possessing significant antioxidant and anti microbial activities.

Study of the Antimicrobial Activity of Aminoreductone against Pathogenic Bacteria in Comparison with Other Antibiotics

Antimicrobial activities of aminoreductone (AR), a product formed in the initial stage of Maillard reaction, were screened against pathogenic bacteria. A significant growth inhibition of AR against all 7 isolates (Staphylococcus aureus ATCC® 25923™, Salmonella typhimurium ATCC® 14028™, Bacillus cereus ATCC® 13061™, Bacillus subtilis ATCC® 11774™, Escherichia coli ATCC® 25922™, Enterococcus faecalis ATCC® 29212™, Listeria innocua ATCC® 33090™) were observed by the standard disc diffusion methods. The inhibition zone for each isolate by AR (2.5 mg) ranged from 15±0mm to 28.3±0.4mm in diameter. The minimum inhibitory concentration (MIC) of AR ranging from 20mM to 26mM was proven in the 7 isolates tested. AR also showed the similar effect of growth inhibition in comparison with antibiotics frequently used for the treatment of infections bacteria, such as amikacin, ciprofloxacin, meropennem and levofloxacin. The results indicated that foods containing AR are valuable sources of bioactive compounds towards pathogenic bacteria.

Leaching of Mineral Nitrogen and Phosphate from Rhizosphere Soil Stressed by Drought and Intensive Rainfall

This work presents the first results from the long-term experiment, which is focused on the impact of intensive rainfall and long period of drought on microbial activities in soil. Fifteen lysimeters were prepared in the area of our interest. This area is a protection zone of underground source of drinking water. These lysimeters were filed with topsoil and subsoil collected in this area and divided into two groups. These groups differ in fertilization and amount of water received during the growing season. Amount of microbial biomass and leaching of mineral nitrogen and phosphates were chosen as main indicators of microbial activities in soil. Content of mineral nitrogen and phosphates was measured in soil solution, which was collected from each lysimeters. Amount of microbial biomass was determined in soil samples that were taken from the lysimeters before and after the long period of drought and intensive rainfall.

Characterization of Antioxidant Peptides of Soybean Protein Hydrolysate

In order to characterize the soy protein hydrolysate obtained in this study, gel chromatography on Sephadex G-25 was used to perform the separation of the peptide mixture and electrophoresis in SDS-polyacrylamide gel has been employed. Protein hydrolysate gave high antioxidant activities, but didn't give any antimicrobial activities. The antioxidant activities of protein hydrolysate was in the same trend of peptide content which gave high antioxidant activities and high peptide content between fractions 15 to 50. With increasing peptide concentrations, the scavenging effect on DPPH radical increased until about 70%, thereafter reaching a plateau. In compare to different concentrations of BHA, which exhibited higher activity (90%), soybean protein hydrolysate exhibited high antioxidant activities (70%) at a concentration of 1.45 mg/ml at fraction 25. Electrophoresis analysis indicated that, low- MW hydrolysate fractions (F1) appeared, on average, to have higher DPPH scavenging activities than high-MW fractions. These results revealed that soybean peptides probably contain substances that were proton donors and could react with free radicals to convert them to stable diamagnetic molecules. 

Antibacterial Activity of the Chennopodium album Leaves and Flowers Extract

Recent years have instance that there is a invigoration of interest in drug discovery from medicinal plants for the support of health in all parts of the world . This study was designed to examine the in vitro antimicrobial activities of the flowers and leaves methanolic and ethanolic extracts of Chenopodium album L. Chenopodium album Linn. flowers and leaves were collected from East Esfahan, Iran. The effects of methanolic and ethanolic extracts were tested against 4 bacterial strains by using disc,well-diffusion method. Results showed that flowers and leaves methanolic and ethanolic extracts of C.album don-t have any activity against the selected bacterial strains. Our study has indicated that ,there are effective different factors on antimicrobial properties of plant extracts

Removal of Cationic Heavy Metal and HOC from Soil-Washed Water Using Activated Carbon

Soil washing process with a surfactant solution is a potential technology for the rapid removal of hydrophobic organic compound (HOC) from soil. However, large amount of washed water would be produced during operation and this should be treated effectively by proper methods. The soil washed water for complex contaminated site with HOC and heavy metals might contain high amount of pollutants such as HOC and heavy metals as well as used surfactant. The heavy metals in the soil washed water have toxic effects on microbial activities thus these should be removed from the washed water before proceeding to a biological waste-water treatment system. Moreover, the used surfactant solutions are necessary to be recovered for reducing the soil washing operation cost. In order to simultaneously remove the heavy metals and HOC from soil-washed water, activated carbon (AC) was used in the present study. In an anionic-nonionic surfactant mixed solution, the Cd(II) and phenanthrene (PHE) were effectively removed by adsorption on activated carbon. The removal efficiency for Cd(II) was increased from 0.027 mmol-Cd/g-AC to 0.142 mmol-Cd/g-AC as the mole ratio of SDS increased in the presence of PHE. The adsorptive capacity of PHE was also increased according to the SDS mole ratio due to the decrement of molar solubilization ratios (MSR) for PHE in an anionic-nonionic surfactant mixture. The simultaneous adsorption of HOC and cationic heavy metals using activated carbon could be a useful method for surfactant recovery and the reduction of heavy metal toxicity in a surfactant-enhanced soil washing process.

The Effect of Dispersed MWCNTs Using SDBS Surfactant on Bacterial Growth

Carbon nanotubes (CNTs) are attractive because of their excellent chemical durability mechanical strength and electrical properties. Therefore there is interest in CNTs for not only electrical and mechanical application, but also biological and medical application. In this study, the dispersion power of surfactant-treated multiwalled carbon nanotubes (MWCNTs) and their effect on the antibacterial activity were examined. Surfactant was used sodium dodecyl-benzenesulfonate (SDBS). UV-vis absorbance and transmission electron microscopy(TEM) were used to characterize the dispersion of MWCNTs in the aqueous phase, showing that the surfactant molecules had been adsorbed onto the MWCNTs surface. The surfactant-treated MWCNTs exhibited antimicrobial activities to streptococcus mutans. The optical density growth curves and viable cell number determined by the plating method suggested that the antimicrobial activity of surfactant-treated MWCNTs was both concentration and treatment time-dependent.

Biological Effects of a Carbohydrate-Binding Protein from an Annelid, Perinereis nuntia Against Human and Phytopathogenic Microorganisms

Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

In Vitro Antibacterial and Antifungal Effects of a 30 kDa D-Galactoside-Specific Lectin from the Demosponge, Halichondria okadai

The present study has been taken to explore the screening of in vitro antimicrobial activities of D-galactose-binding sponge lectin (HOL-30). HOL-30 was purified from the marine demosponge Halichondria okadai by affinity chromatography. The molecular mass of the lectin was determined to be 30 kDa with a single polypeptide by SDS-PAGE under non-reducing and reducing conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed rabbit and human erythrocytes with preference for type O erythrocytes. The lectin was subjected to evaluation for inhibition of microbial growth by the disc diffusion method against eleven human pathogenic gram-positive and gram-negative bacteria. The lectin exhibited strong antibacterial activity against gram-positive bacteria, such as Bacillus megaterium and Bacillus subtilis. However, it did not affect against gram-negative bacteria such as Salmonella typhi and Escherichia coli. The largest zone of inhibition was recorded of Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in diameter) at a concentration of the lectin (250 μg/disc). On the other hand, the antifungal activity of the lectin was investigated against six phytopathogenic fungi based on food poisoning technique. The lectin has shown maximum inhibition (22.83%) of mycelial growth of Botrydiplodia theobromae at a concentration of 100 μg/mL media. These findings indicate that the lectin may be of importance to clinical microbiology and have therapeutic applications.