Search results for: rhizopus
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31

Search results for: rhizopus

31 Improving the Quality of Casava Peel-Leaf Mixture through Fermentation with Rhizopus oligosporusas Poultry Ration

Authors: Mirnawati, G. Ciptaan, Ferawati

Abstract:

This study aims to improve the quality of the cassava peel-leaf mixture (CPLM) through fermentation with Rhizopus oligosporusas poultry ration. This research is an experimental study using a completely randomized design (CRD) with four treatments and five replications. The treatments were cassava peel-leaf mixture (CPLM) fermented with Rhizopus oligosporus. The treatments were a combination of cassava peel and leaves with the ratio of; A (9:1), B (8:2), C (7:3), and D (6:4). The observed variables were protease enzyme activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and metabolic energy. The results of the diversity analysis showed that there was a very significant (p < 0.01) effect on protease activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and energy metabolism of fermented CPLM. Based on the results of the study, it can be concluded that CPLM (6:4) fermented with Rhizopus oligosporus gave the best results seen from protease activity 7,25 U/ml, 21.23% crude protein, 19.80% crude fiber, 59.65% nitrogen retention, 62.99% crude fiber digestibility and metabolic energy 2671 Kcal/kg.

Keywords: quality, Casava peel-leaf mixture, fermentation, Rhizopus oligosporus

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30 Synthesis of DHA Rich Glycerides with Immobilized Lipases from Mucor miehei and Rhizopus oryzae

Authors: Satyendra P. Chaurasia, Aditi Sharma, Ajay K. Dalai

Abstract:

The esterification of Docosahexaenoic acid (DHA) with glycerol using immobilized Mucor mie-hei lipase (MML) and Rhizopus oryzae lipase (ROL) have been studied in the present paper to synthesize triglycerides (TG) rich in DHA. Both immobilized lipases (MML and ROL), and their support materials (immobead-150 and ion-exchange resin) were characterized and compared for surface properties with BET, for chemical functional groups with FT-IR, and for particle size distribution with particle size analyzer. The most suitable reaction conditions for synthesis of DHA rich TG in biphasic solvent system were found as 1:3 (wt/wt) glycerol to DHA ratio, 1:1 (wt/wt) buffer to DHA ratio, 1:1 (wt/wt) solvent to DHA ratio at 50 ºC temperature, and 600 rpm speed of agitation with 100 mg of immobilized lipases. Maximum 95.9 % esterification was obtained with immobilized MML in 14 days reaction with formation of 65.7 wt% DHA rich TG. Whereas, immobilized ROL has shown formation of only 23.8 wt% DHA rich TG with total 78.9 % esterification in 15 days. Additionally, repeated use of both immobilized lipases was con-ducted up to five cycles, indicated 50.4% and 41.2 % activity retention after fifth repeated use of immobilized MML and ROL, respectively.

Keywords: DHA, immobilized Mucor miehei lipase, Rhizopus oryzae lipase, esterification

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29 Biodegradation Potential of Selected Micromycetes against Dyeing Unit Effluents of Sapphire Industry in Raiwind Road Lahore

Authors: Samina Sarwar, Hajra Khalil

Abstract:

Mycoremediation is emerging as a potential approach for eco-friendly and cost-effective remediation of polluted effluents collected from the dyeing unit of the textile industry was examined. This work dealt with the analyses of the bio remedial capability of some potential indigenous six fungal isolates viz., Aspergillus alliaceus, Aspergillus flavus, Aspergillus fumigatus Aspergillus niger, Penicillium sp. and Rhizopus oryzae were identified and selected for studies. All fungal species were known to bring bioremediation, which had been confirmed by measuring the percentage reduction potential in different parameters, i.e., pH, Electrical Conductivity (EC), Total Suspended Solids (TSS), Total Dissolved Solids (TDS), Biological Oxygen Demand (BOD) and Chemical Oxygen Demand (COD). Rhizopus oryzae showed the highest reduction in pH, EC, and BOD, while Aspergillus fumigatus showed the highest reduction in TDS and TSS, and COD under the optimal conditions of this study. The biodegradation potential of these fungal species was confirmed, evidenced by excellent evaluation of experimental data to propose Rhizopus oryzae and Aspergillus fumigatus as a cost-effective solution to treat the effluents from the dyeing unit of the textile industry.

Keywords: biological reduction, fungal isolates, micromycetes, mycoremediation

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28 Biodegradation Potential of Selected Micromycetes Against Dyeing Unit Effluents of Sapphire Industry, Raiwind Road Lahore

Authors: Samina Sarwar, Hajra Khalil

Abstract:

Mycoremediation is emerging as a potential approach for eco-friendly and cost-effective remediation of polluted effluents collected from the dyeing unit of the textile industry was examined. This work dealt with the analyses of the bio remedial capability of some potential indigenous six fungal isolates viz., Aspergillus alliaceus, Aspergillus flavus, Aspergillus fumigatus Aspergillus niger, Penicillium sp. and Rhizopus oryzae were identified and selected for studies. All fungal species were known to bring bioremediation, which had been confirmed by measuring the percentage reduction potential in different parameters, i.e., pH, Electrical Conductivity (EC), Total Suspended Solids (TSS), Total Dissolved Solids (TDS), Biological Oxygen Demand (BOD) and Chemical Oxygen Demand (COD). Rhizopus oryzae showed the highest reduction in pH, EC, and BOD, while Aspergillus fumigatus showed the highest reduction in TDS and TSS, and COD under the optimal conditions of this study. The biodegradation potential of these fungal species was confirmed, evidenced by excellent evaluation of experimental data to propose Rhizopus oryzae and Aspergillus fumigatus as a cost-effective solution to treat the effluents from the dyeing unit of the textile industry.

Keywords: biological reduction, fungal isolates, micromycetes, mycoremediation

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27 Isolation, Identification and Screening of Pectinase Producing Fungi Isolated from Apple (Malus Domestica)

Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar

Abstract:

Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.

Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus

Procedia PDF Downloads 27
26 Bioprotective Role of Soil Borne Bacillus Strains against Selected Fungal Pathogens of Agriculture Relevance

Authors: Asad Ali, Asif Jamal

Abstract:

The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: bacillus species, biocontrol agent, biopesticides, phytopathogens

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25 Bioprotective Role of Soil Borne Bacillus Strain against Selected Fungal Pathogens of Agriculture Relevance

Authors: Asif Jamal, Asad Ali, Muhammad Ishtiaq Ali

Abstract:

The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis, and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as a bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: biological control, Bacillus spp, fungal pathogens, agriculture

Procedia PDF Downloads 239
24 Efficiency for Enzyme Production of Fungi Isolated from the Stomach of Buffalo

Authors: Suphalucksana, Wichai, Sangsoponjit Settasit, Soytong Kasem

Abstract:

A study on the efficiency for enzyme production of fungi isolated from stomach of buffalo was conducted. The fungi were collected from 4 parts of stomach as rumen, reticulum, omasum and abomasums. The objective to study the efficiency of fungi from stomach of buffalo had effected to produced enzyme and to selected fungi for their ability to produced enzyme cellulase, hemicellulase and ligninase. Results shown that the fungi isolated from rumen were: Eupenicillium sp. (B-RU-01-1), Eupenicillium sp. (B-RU-02-3G), Rhyzopus stolonifer (B-RU-01-4) and Trichoderma sp. (B-RU-01-2). From the reticulum, Aspergillus glaucus (B-RET-02-3), Aspergillus orchraceus (B-RET-02-2) and Penicillium sp. (B-RET-02-4) were found. In the omasum Aspergillus fumigatus (B-OMA-01-1G), Eurotium sp. (B-OMA-01-4) and Rhizopus stolonifer (B-OMA-02-3) were isolated and in the abomasums Aspergillus flavas (B-ABO-02-3), Aspergillus fumigatus (B-ABO-02-1), Aspergillus niger (B-ABO-01-3G), Aspergillius terreus (B-ABO-02-4) and Mucor sp. (B-ABO-02-4G). Results of enzyme analysis revealed that cellulase was produced by isolated: Eupenicillium sp. (B-RU-02-3G), Eupenicillium sp. (B-RU-01-1), Penicillium sp. (B-RET-02-4), Aspergillius glaucus (B-RET-02-3), Aspergillus ochraceus (B-RET-02-2), Aspergillius fumigatus (B-OMA-01-1G), Eurotium sp. (B-OMA-01-4), Aspergillius flavus (B-ABO-02-3), Aspergillius fumigatus (B-ABO-02-1), Aspergillius niger (B-ABO-01-3G), Aspergillius terreus (B-ABO-02-4). Hemicellulase was produced Eupenicillium sp. (B-RU-02-3G), Eupenicillium sp. (B-RU-01-1), Rhizopus stolonifer (B-RU-01-4), Trichoderma sp. (B-RU-01-2), Aspergillius glaucus (B-RET-02-3), Aspergillus ochraceus (B-RET-02-2), Penicillium sp. (B-RET-02-4), Aspergillius fumigatus (B-OMA-01-1G), Eurotium sp. (B-OMA -01-4), Aspergillius flavus (B-ABO-02-3), Aspergillius fumigatus (B-ABO-02-1) Aspergillius niger (B-ABO-01-3G), Aspergillius terreus (B-ABO-02-4), Mucor sp. (B-ABO-02-4G). For the enzyme ligninase, two isolates were found to produced this enzyme namely : Trichoderma sp. (B-RU-01-2) and Mucor sp. (B-ABO-02-4G).

Keywords: enzyme production from fungi, enzyme production, fungi, agricultural technology

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23 Proximate Composition and Sensory Properties of Complementary Food from Fermented Acha (Digitaria exilis), Soybean and Orange-Flesh Sweet Potato Blends

Authors: N. C. Okoronkwo, I. E. Mbaeyi-Nwaoha, C. P. Agbata

Abstract:

Childhood malnutrition is one of the most persistent public health problems throughout developing countries, including Nigeria. Demographic and Health survey data from twenty-one developing countries indicated that poor complementary feeding of children aged 6- 23 months contributes to negative growth trends. To reduce malnutrition among children in the society, formulation of complimentary food rich in essential nutrient for optimum growth and development of infants is essential. This study focused on the evaluation of complementary food produced by solid-state fermentation of Acha and Soybean using Rhizopus oligosporus (2710) and Orange-fleshed sweet potatoes (OFSP) using Lactobacillus planterum (B-41621). The raw materials were soaked separately, each in four volumes of 0.9M acetic acid for 16 hours, rinsed with clean water, steam cooked and cooled. Solid-state fermentation (SSF) was carried out by inoculating Acha and Soybean with spore suspension (1x 10⁶spores/ml) of Rhizopus oligosporus (2710) and OFSP with spore suspension (1x 106spores/ml) of Lactobacillus planterum (B-41621). Fermentation which lasted for 72hours was carried out with 24hours sampling. The samples were blended in the following ratios: Acha and soybean 100: 100 (AS), Acha/soybean and OFSP 50: 50(ASO), made into gruel and compared with a commercial infant formula (Cerelac) which served as the control (CTRL). The samples were analyzed for proximate composition using AOAC methods and sensory attributes using a hedonic scale. Results showed that moisture, crude protein, fibre and ash content increased significantly (p<0.05) as fermentation progressed, while carbohydrate and fat content decreased. The protein, moisture, fibre and ash content ranged from 17.10-19.02%, 54.97-56.27%, 7.08-7.60% and2.09-2.38%, respectively, while carbohydrate and fat content ranged from 12.95-10.21% and 5.81-4.52%, respectively. In sensory scores, there were no significant (p>0.05) difference between the average mean scores of colours, texture and consistency of the samples. The sensory score for the overall acceptability ranged from 6.20-7.80. Sample CTRL had the highest score, while sample ASO had the least score. There was no significant (p>0.05) difference between samples CTRL and AS. Solid-state fermentation improved the nutritional content and flavour of the developed complementary food, which is needed for infant growth and development.

Keywords: Complementary food, malnutrition, proximate composition, solid-state fermentation

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22 Control of Fungal Growth in Sweet Orange and Mango Juices by Justica flava and Afromomum melegueta Extracts

Authors: Adferotimi Banso

Abstract:

A laboratory investigation was conducted to determine the effect of Justica flava and Aframonium melegueta on the growth of Aspergillus niger, Rhizopus stolonifer and Fusarium species in sweet orange and mango juices. Aqueous extract (3%v/v) of Justica flava and Aframonium melegueta reduced the growth of the fungi, a combination of 2% (v/v) each of Justica flava and Aframonium melegueta extracts reduced the growth better. Partial purification of aqueous extracts of Justica flava and Aframonium melegueta showed that ethyl acetate fraction of the extracts exhibited the highest level of inhibition of growth of the test fungi compared with diethyl ether and n-hexane fractions. The results suggest that extracts of Justica flava and Aframonium melegueta may be important substitutes for conventional chemical preservatives in the processing of fruit juices.

Keywords: aqueous, fraction, mango, orange, purification, sweet

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21 Antagonist Study of Fungi Isolated from the Burned Forests of Region of Mila, Algeria

Authors: Abdelaziz Wided, Khiat Nawel, Khiat Inssaf

Abstract:

The present study was initiated to: Determine burned forest-inhabiting fungi in Zouagha, Terri Beinène, Mila and study the antagonistic activity of Trichoderma sp against Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp. 18 fungal strains were isolated from Soil samples taken from the forest Zouagha (Burned) in the region Mila representing 6 genera: Trichoderma sp et Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp, Rhizopus sp. The tests of dual culture method on culture medium (PDA) against Trichoderma sp et Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp revealed that: Trichoderma sp could reduce l mycelium grouth of Fusarium sp23.13%, Penicillium sp33.13%, Rhizoctoniasp33.75 %and Alternaria sp 38.31% in comparaison with the witness after 6 days at room temperature. The strains of Fusarium sp ,Penicillium sp, Rhizoctonia sp et Alternaria sp showed differences sensibility to the antagoniste.

Keywords: isolation, identification, molds, burned soil of zouagha, antagonism, trichoderma sp

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20 The Effect of Some Microorganisms from Gastrointestinal Tracts on the Nutritive Value of Broiler Diets

Authors: S. Sangsoponjit, W. Suphalucksana, K. Soytong

Abstract:

A 2x2 factorial experiment was carried out to determine the effects of two levels of diet supplemented with and without microorganisms in combination with and without feed sterilisation on the nutritive value of broiler diets with four replications in each treatment. Some microorganisms from the gastrointestinal tracts of chickens were supplemented in commercial broiler diets. They were bacterial (BC-NA-01), actinomycetes (BI-NA-03, BC-NA-02 and BL-NA-02), Aspergillus niger sp.(BD-PDA-01), Mucor sp.(BL-PDA-02), Rhizopus stolonifer sp.(BI-PDA-02) and Trichoderma sp.(BL-PDA-02). The results of the proximate analysis revealed that the diet supplemented with microorganisms had a higher percentage of DM and CF in the starter diet(0-3 wks), grower diet(4-5wks) and finisher diet (last period) than the diet without microorganisms (p<0.05). Also, they were higher in the percentage of CP in the starter diet and EE in both the starter diet and grower diet than the diet without microorganisms (p<0.05). The sterilised diet had a higher percentage of moisture than the non-sterilized diet (p<0.01). Also, they were higher in the percentage of CP in the starter diet and CF in both the grower diet and finisher diet than the non-sterilized diet (p<0.05). The sterilized diet supplemented with microorganisms was higher in ME than the non-sterilize diet without microorganisms in the starter diet, grower diet and finisher diet (P<0.01).

Keywords: microorganisms, gastrointestinal tract, nutritive value, broiler diets

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19 Antifungal Protein ~35kDa Produced by Bacillus cereus Inhibits the Growth of Some Molds and Yeasts

Authors: Saleh H. Salmen, Sulaiman Ali Alharbi, Hany M. Yehia, Mohammad A. Khiyami, Milton Wainwright, Naiyf S. Alharbi, Arunachalam Chinnathambi

Abstract:

An antifungal protein synthesized by Bacillus cereus has been partially purified by the use of ammonium sulfate precipitation and Sephadex-G-200 column chromatography. The protein was produced from Bacillus cereus grown in potato Dextrose Broth Medium (PDB) at 30 ºC for 3 days at 100 rpm. The protein showed antagonistic effect against some fungi and yeasts. Crude extract from medium and semi-purified protein were tested in vitro against both fungi and yeasts using the disc diffusion method in order to detect the inhibitory effect of the protein. Zones of inhibition of the following diameter were found (mm) were Alternaria alternate (28), Rhodotorula glutinis (20), Fusarium sp. (16), Rhizopus sp. (15), Penicillium digitatum (13), Mucor sp. (13) and Aspergillus niger (10). The isolated protein was found to have a molecular weight of ~35kDa by sodium deodecyl sulfate-poly acrylamide gel electrophoresis. The data showed that the protein of Bacillus cereus has antifungal activity, a fact which points to the possibility of using it as a bio-control agent against some fungi, findings which emphasize the potential role of B. cereus as an important bio-control agent.

Keywords: bacillus cereus, ~35kDa protein, molds, yeasts

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18 Impact of Calcium Carbide Waste Dumpsites on Soil Chemical and Microbial Characteristics

Authors: C. E. Ihejirika, M. I. Nwachukwu, R. F. Njoku-Tony, O. C. Ihejirika, U. O. Enwereuzoh, E. O. Imo, D. C. Ashiegbu

Abstract:

Disposal of industrial solid wastes in the environment is a major environmental challenge. This study investigated the effects of calcium carbide waste dumpsites on soil quality. Soil samples were collected with hand auger from three different dumpsites at varying depths and made into composite samples. Samples were subjected to standard analytical procedures. pH varied from 10.38 to 8.28, nitrate from 5.6mg/kg to 9.3mg/kg, phosphate from 8.8mg/kg to 12.3mg/kg, calcium carbide reduced from 10% to to 3%. Calcium carbide was absent in control soil samples. Bacterial counts from dumpsites ranged from 1.8 x 105cfu/g - 2.5 x 105cfu/g while fungal ranged from 0.8 x 103cfu/g - 1.4 x 103cfu/g. Bacterial isolates included Pseudomonas spp, Flavobacterium spp, and Achromobacter spp, while fungal isolates include Penicillium notatum, Aspergillus niger, and Rhizopus stolonifer. No organism was isolated from the dumpsites at soil depth of 0-15 cm, while there were isolates from other soil depths. Toxicity might be due to alkaline condition of the dumpsite. Calcium carbide might be bactericidal and fungicidal leading to cellular physiology, growth retardation, death, general loss of biodiversity and reduction of ecosystem processes. Detoxification of calcium carbide waste before disposal on soil might be the best option in management.

Keywords: biodiversity, calcium-carbide, denitrification, toxicity

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17 Storage of Maize Grains Using Powder and Oils of Commonly Used Medicinal Plants (Aframomum melegueta, Garcinia kola and Piper guineense)

Authors: T. O. Adejumo, O. S. Akinyemi

Abstract:

Powders and oils of Aframomum melegueta, Garcinia kola and Piper guineense were tested as preservatives for the storage of maize grains for four weeks. The concentrations of the powders were 32.5gkg-1, 65.0gkg-1 and 97.5gkg-1 maize, while those of oils were 0.85mlkg-1, 0.50mlkg-1 and 0.75mlkg-1 maize respectively. Powders of the three botanicals at 97.5gkg-1 maize possessed insecticidal effect on Sitophilus zeamais and also inhibitory activities on Aspergillus flavus, A. fumigatus, A. niger and Fusarium verticillioides, while little effect was observed for other concentrations. Oils of the three botanicals at 0.50mlkg-1 and 0.75mlkg-1 maize showed an insecticidal effect on S. zeamais and also inhibitory activities on A. flavus, A. fumigatus, A. niger, F. verticillioides, Penicillium and Rhizopus species. Oils showed more potential as a protectant against fungal and insect pest in storage maize grains than powders. Powders and oils of A. melegueta, G. kola and P. guineense could be successfully used as biopesticides.

Keywords: aframomum melegueta, garcinia kola, maize, powder, oils, piper guineense

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16 Encapsulation of Satureja khuzestanica Essential Oil in Chitosan Nanoparticles with Enhanced Antifungal Activity

Authors: Amir Amiri, Naghmeh Morakabati

Abstract:

During the recent years the six-fold growth of cancer in Iran has led the production of healthy products to become a challenge in the food industry. Due to the young population in the country, the consumption of fast foods is growing. The chemical cancer-causing preservatives are used to produce these products more than the standard; so using an appropriate alternative seems to be important. On the one hand, the plant essential oils show the high antimicrobial potential against pathogenic and spoilage microorganisms and on the other hand they are highly volatile and decomposed under the processing conditions. The study aims to produce the loaded chitosan nanoparticles with different concentrations of savory essential oil to improve the anti-microbial property and increase the resistance of essential oil to oxygen and heat. The encapsulation efficiency was obtained in the range of 32.07% to 39.93% and the particle size distribution of the samples was observed in the range of 159 to 210 nm. The range of Zeta potential was obtained between -11.9 to -23.1 mV. The essential oil loaded in chitosan showed stronger antifungal activity against Rhizopus stolonifer. The results showed that the antioxidant property is directly related to the concentration of loaded essential oil so that the antioxidant property increases by increasing the concentration of essential oil. In general, it seems that the savory essential oil loaded in chitosan particles can be used as a food processor.

Keywords: chitosan, encapsulation, essential oil, nanogel

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15 Associated Mycoflora AF Mucuna Sloanei Seeds and Their Effects on Nutritional and Phytochemical Contents of the Seeds

Authors: U.N. Emiri, E. Moroyei

Abstract:

Mycoflora associated with the seed rot disease of Mucuna sloanei and their effects on nutrient and phytochemical composition of the seeds were investigated. The fungal pathogens implicated in the seed rot disease were Rhizopus stolonifer, Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum. The fungal isolates were aseptically inoculated into healthy M. Sloanei seeds and incubated for 7 days at room temperature of 25 ± 30c. The results of the proximate and mineral analysis in mg/100g of fungal infected and non-infected (control) seeds that were carried out revealed that there was an increase in Moisture and Carbohydrate content of the fungal infected seeds relative to the non-infected seeds (control). However, there was a decrease in Ash, Fibre, Lipid, and Protein content of the fungal infected seeds relative to the non-infected (control). It was observed that moisture had increased from 10.50 ± 0.16 in the non-infected seeds to 17.60 ± 0.20 in the infected samples and Carbohydrate content had also increased from 49.6 ± 0.25 in the non-infected to 52.50 ± 0.29 in the infected seeds. The following parameters decreased in the infected than in the non-infected seeds. They include Ash 2.60 ± 0.12, Crude fibre 1.9 ± 0.08, Lipid 6.50 ± 0.16, and Protein content 18.50 ± 0.06. Similarly, Calcium 2.50 ± 0.12, Phosphorus 1.80 + 0.12 and Potassium 1.80 + 0.09 increased in the infected than in the non-infected seed, while iron 0.20 ± 0.05, Sodium 0.02 ± 0.01 and Magnesium 0.06 ± 0.02 decreased in the infected seeds. All phytochemical contents analyzed increased in the infected seeds viz Tannim 0.50 ± 0.12, Oxalate 1.60 ± 0.05, Hydrogen cyanide 1.82 ± 0.06, and Saponin 2.50+0.28. However, the nutrient compositions and Phytochemical between the infected and non-infected seeds are not significantly different (p > 0.05).

Keywords: Mycoflora, mucuna sloanei, seeds, phytochemical, nutrient composition

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14 Treatment of Grey Water from Different Restaurants in FUTA Using Fungi

Authors: F. A. Ogundolie, F. Okogue, D. V. Adegunloye

Abstract:

Greywater samples were obtained from three restaurants in the Federal University of Technology; Akure coded SSR, MGR and GGR. Fungi isolates obtained include Rhizopus stolonifer, Aspergillus niger, Mucor mucedo, Aspergillus flavus, Saccharomyces cerevisiae. Of these fungi isolates obtained, R. stolonifer, A. niger and A. flavus showed significant degradation ability on grey water and was used for this research. A simple bioreactor was constructed using biodegradation process in purification of waste water samples. Waste water undergoes primary treatment; secondary treatment involves the introduction of the isolated organisms into the waste water sample and the tertiary treatment which involved the use of filter candle and the sand bed filtration process to achieve the end product without the use of chemicals. A. niger brought about significant reduction in both the bacterial load and the fungi load of the greywater samples of the three respective restaurants with a reduction of (1.29 × 108 to 1.57 × 102 cfu/ml; 1.04 × 108 to 1.12 × 102 cfu/ml and 1.72 × 108 to 1.60 × 102 cfu/ml) for bacterial load in SSR, MGR and GGR respectively. Reduction of 2.01 × 104 to 1.2 × 101; 1.72 × 104 to 1.1 × 101, and 2.50 × 104 to 1.5 × 101 in fungi load from SSR, MGR and GGR respectively. Result of degradation of these selected waste water by the fungi showed that A. niger was probably more potent in the degradation of organic matter and hence, A. niger could be used in the treatment of wastewater.

Keywords: Aspergillus niger, greywater, bacterial, fungi, microbial load, bioreactor, biodegradation, purification, organic matter and filtration

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13 Effects of Storage Methods on Proximate Compositions of African Yam Bean (Sphenostylis stenocarpa) Seeds

Authors: Iyabode A. Kehinde, Temitope A. Oyedele, Clement G. Afolabi

Abstract:

One of the limitations of African yam bean (AYB) (Sphenostylis sternocarpa) is poor storage ability due to the adverse effect of seed-borne fungi. This study was conducted to examine the effects of storage methods on the nutritive composition of AYB seeds stored in three types of storage materials viz; Jute bags, Polypropylene bags, and Plastic Bowls. Freshly harvested seeds of AYB seeds were stored in all the storage materials for 6 months using 2 × 3 factorial (2 AYB cultivars and 3 storage methods) in 3 replicates. The proximate analysis of the stored AYB seeds was carried out at 3 and 6 months after storage using standard methods. The temperature and relative humidity of the storeroom was recorded monthly with Kestrel pocket weather tracker 4000. Seeds stored in jute bags gave the best values for crude protein (24.87%), ash (5.69%) and fat content (6.64%) but recorded least values for crude fibre (2.55%), carbohydrate (50.86%) and moisture content (12.68%) at the 6th month of storage. The temperature of the storeroom decreased from 32.9ºC - 28.3ºC, while the relative humidity increased from 78% - 86%. Decreased incidence of field fungi namely: Rhizopus oryzae, Aspergillus flavus, Geotricum candidum, Aspergillus fumigatus and Mucor meihei was accompanied by the increase in storage fungi viz: Apergillus niger, Mucor hiemalis, Penicillium espansum and Penicillium atrovenetum with prolonged storage. The study showed that of the three storage materials jute bag was more effective at preserving AYB seeds.

Keywords: storage methods, proximate composition, African Yam Bean, fungi

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12 Prospects in Development of Ecofriendly Biopesticides in Management of Postharvest Fungal Deterioration of Cassava (Manihot esculenta Crantz)

Authors: Anderson Chidi Amadioha, Promise Chidi Kenkwo, A. A. Markson

Abstract:

Cassava (Manihot esculenta Crantz) is an important food and cash crop that provide cheap source of carbohydrate for food, feed and raw material for industries hence a commodity for feature economic development of developing countries. Despite the importance, its production potentials is undermined by disease agents that greatly reduce yield and render it unfit for human consumption and industrial use. Pathogenicity tests on fungal isolates from infected cassava revealed Aspergillus flavus, Rhizopus stolonifer, Aspergillus niger, and Trichodderma viride as rot-causing organisms. Water and ethanol extracts of Piper guineense, Ocimum graticimum, Cassia alata, and Tagetes erecta at 50% concentration significantly inhibited the radial growth of the pathogens in vitro and their development and spread in vivo. Low cassava rot incidence and severity was recorded when the extracts were applied before than after spray inoculating with spore suspension (1x105 spores/ml of distilled water) of the pathogenic organisms. The plant materials are readily available, and their extracts are biodegradable and cost effective. The fungitoxic potentials of extracts of these plant materials could be exploited as potent biopesticides in the management of postharvest fungal deterioration of cassava especially in developing countries where synthetic fungicides are not only scarce but also expensive for resource poor farmers who produce over 95% of the food consumed.

Keywords: cassava, biopesticides, in vitro, in vivo, pathogens, plant extracts

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11 Cytotoxic Activity of Acetone and Ethanol Overripe Tempe Extracts against MCF-7 Breast Cancer Cells and Their Antioxidant Property

Authors: Dian Muzdalifah, Anastasia F. Devi, Zatil A. Athaillah, Linar Z. Udin

Abstract:

Tempe is a functional food prepared from soybeans through Rhizopus spp fermentation. It is well known as functional food, originated from Indonesia. Most studies on tempe functionalities refer to ripe (48 h fermentation) tempe and only limited studies discuss overripe tempe while longer fermentation time possibly increased tempe health benefit. Hence, the present study was performed to investigate the cytotoxic activity againts MCF-7 breast cancer cells and antioxidant property of tempe prepared from 0–156 h of fermentation. Tempe samples were dried and extracted with acetone and ethanol, respectively. Their extracts were used for subsequent analysis. The cytotoxic activity was assessed on MCF 7 breast cancer cells using Alamar Blue method. The antioxidant activity was determined by DPPH free radical scavenging assay. The results indicated that acetone extracts of 108 h tempe had a potent cytotoxic activity against MCF-7 breast cancer cells (IC50 = 2.54 ± 0,30 μg/mL). Ethanol extracts of 108 h tempe also showed the potency, but at slightly higher IC50 (5.20 ± 1.01 μg/mL). Both acetone and ethanol extracts of 108 and 120 h tempe showed high antioxidant activity expressed as percent inhibition with no significant difference. However, acetone extracts of 120 h tempe (81.31 ± 3.70 %) had better ability to inhibit oxidation reaction than that of ethanol extracts (75.77 ± 6.00 %). It can be concluded that the cytotoxic activity of tempe from 0–156 h of fermentation is positively correlated to their corresponding antioxidant property. Longer fermentation time, up to 108 h, increased the ability of tempe to inhibit the growth of MCF-7 breast cancer cells and oxidative reaction. But extended fermentation time, up to 156 h, tends to decrease its ability. Further studies are encouraged to identify the active components contained in each extract.

Keywords: antioxidant property, cytotoxic activity, extracts, overripe tempeh

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10 Ethanol and Biomass Production from Spent Sulfite Liquor by Filamentous Fungi

Authors: M. T. Asadollahzadeh, A. Ghasemian, A. R. Saraeian, H. Resalati, P. R. Lennartsson, M. J. Taherzadeh

Abstract:

Since filamentous fungi are capable of assimilating several types of sugars (hexoses and pentoses), they are potential candidates for bioconversion of spent sulfite liquor (SSL). Three filamentous fungi such as Aspergillus oryzae, Mucor indicus, and Rhizopus oryzae were investigated in this work. The SSL was diluted in order to obtain concentrations of 50, 60, 70, 80, and 90% and supplemented with two types of nutrients. The results from cultivations in shake flask showed that A. oryzae and M. indicus were not able to grow in pure SSL and SSL90% while R. oryzae could grow only in SSL50% and SSL60%. Cultivation with A. oryzae resulted in the highest yield of produced fungal biomass, while R. oryzae cultivation resulted in the lowest fungal biomass yield. Although, the mediums containing yeast extract, (NH4)2SO4, KH2PO4, CaCl2∙2H2O, and MgSO4∙7H2O as nutrients supplementations produced higher fungal biomass compared to the mediums containing NH4H2PO4 and ammonia, but there was no significant difference between two types of nutrients in terms of sugars and acetic acid consumption rate. The sugars consumption in M. indicus cultivation was faster than A. oryzae and R. oryzae cultivation. Acetic acid present in SSL was completely consumed during cultivation of all fungi. M. indicus was the best and fastest ethanol producer from SSL among the fungi examined, when yeast extract and salts were used as nutrients supplementations. Furthermore, no further improvement in ethanol concentration and rate of sugars consumption was obtained in medium supplemented with NH4H2PO4 and ammonia compared to medium containing yeast extract, (NH4)2SO4, KH2PO4, CaCl2∙2H2O, and MgSO4∙7H2O. On the other hand, the higher dilution of SSL resulted in a better fermentability, and better consumption of sugars and acetic acid.

Keywords: ethanol, filamentous fungi, fungal biomass, spent sulfite liquor

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9 Fungi Associated with Decline of Kikar (Acacia nilotica) and Red River Gum (Eucalyptus camaldulensis) in Faisalabad

Authors: I. Ahmad, A. Hannan, S. Ahmad, M. Asif, M. F. Nawaz, M. A. Tanvir, M. F. Azhar

Abstract:

During this research, a comprehensive survey of tree growing areas of Faisalabad district of Pakistan was conducted to observe the symptoms, spectrum, occurrence and severity of A. nilotica and E. camaldulensis decline. Objective of current research was to investigate specific fungal pathogens involved in decline of A. nilotica and E. camaldulensis. For this purpose, infected roots, bark, neck portion, stem, branches, leaves and infected soils were collected to identify associated fungi. Potato dextrose agar (PDA) and Czepak dox agar media were used for isolations. Identification of isolated fungi was done microscopically and different fungi were identified. During survey of urban locations of Faisalabad, disease incidence on Kikar and Eucalyptus was recorded as 3.9-7.9% and 2.6-7.1% respectively. Survey of Agroforest zones of Faisalabad revealed decline incidence on kikar 7.5% from Sargodha road while on Satiana and Jhang road it was not planted. In eucalyptus trees, 4%, 8% and 0% disease incidence was observed on Jhang road, Sargodha road and Satiana road respectively. The maximum fungus isolated from the kikar tree was Drechslera australiensis (5.00%) from the stem part. Aspergillus flavus also gave the maximum value of (3.05%) from the bark. Alternaria alternata gave the maximum value of (2.05%) from leaves. Rhizopus and Mucor spp. were recorded minimum as compared to the Drechslera, Alternaria and Aspergillus. The maximum fungus isolated from the Eucalyptus tree was Armillaria luteobubalina (5.00%) from the stem part. The other fungi isolated were Macrophamina phaseolina and A. niger.

Keywords: decline, frequency of mycoflora, A. nilotica and E. camaldulensis, Drechslera australiensis, Armillaria luteobubalina

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8 Amylase Activities of Mould Isolated from Spoilt Ogi and Eko: Two (2) Fermented Maize Products

Authors: Gafar Bamigbade, Adebunkola Omemu

Abstract:

“Ogi” is a fermented cereal gruel prepared from maize (Zea mays), millet (Pennisetum typhoideum) or guinea corn (Sorghum bicolour). It could be boiled to give a thicker consistency wrapped in leaf allowed to cool and set to a gel known as “eko”. The objective of this study is to determine the amylase activities of mould associated with the spoilage of Ogi and eko. Moulds were isolated from spoilt Ogi and eko samples using standard microbiological procedures. The isolate was then screened for amylase production using starch agar medium. Positive isolates were used for amylase production by solid state fermentation (SFF) using rice bran as the medium. An alpha-amylase and glucoamylase activity of the crude enzyme was determined using the DNS method. The mean mold Population ranged from 1.15 X 105cfu/g for raw Ogi to 6.25 X 105cfu/g for Eko (wrapped in Leaves). Twenty-seven (27) moulds isolated from the sample include A. niger, A. flavus, A. fumigatus, Rhizopus species and Penicillium species. Aspergillus flavus had the highest percentage (51.9%) of incidence while Penicillium species had the least (3.7%). Out of the 27 isolates screened, 19 were found to be amylase positive by showing a clear zone around their colony after flooding with iodine solution. Diameter of clear zone ranged from 3.00mm (Aspergillus niger, C4) to 22.00mm (Aspergillus flavus, A1). Aspergillus niger isolated from spoilt Eko wrapped in leaf has the highest percentage alpha-amylase activity (30.8%) and Aspergillus flavus isolated from spoilt raw ogi has the lowest activity (11.4%). Aspergillus niger isolated from spoilt Eko wrapped in nylon produces the highest glucoamylase activity (240U/ml) while penicillium specie isolated from spoilt cooked ogi has the lowest activity (100U/ml). This study shows that moulds associated with spoilage of ogi and eko can produce amylase.

Keywords: glucoamylase, alpha amylase, ogi, eko

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7 Preparation of Tempeh Spores Powder

Authors: Jaruwan Chutrtong, Tanakwan Bussabun

Abstract:

Study production of tempeh inoculums powder by freeze-drying comparison with dry at 50°C and the sun bask for developing efficient tempeh inoculums for tempeh producing. Rhizopus oligosporus in PDA slant cultures was incubated at 30°C for 3-5 days until spores and mycelium. Preparation spores suspension with sterilized water and then count the number of started spores. Fill spores suspension in Rice flour and soy flour, mixed with water (in the ratio 10: 7), which is steamed and sterilized at 121°C 15min. Incubated at room temperature for 4 days, count number of spores. Then take the progressive infection and full spore dough to dry at 50°C, sun bask, and lyophilize. Grind to powder. Then pack in plastic bags, stored at 5°C. To investigate quality of inoculums which use different methods, tempeh was fermented every 4 weeks for 24 weeks of the experiment. The result found that rice flour is not suitable to use as raw material in the production of powdered spores. Fungi can growth rarely. Less number of spores and requires more time than soy flour. For drying method, lyophilization is the least possible time. Samples from this method are very hard and very dark and harder to grind than other methods. Drying at 50°C takes longer time than lyophilization but can also set time use for drying. Character of the dry samples is hard solid and brown color, but can be grinded easier. The sun drying takes the longest time, can’t determine the exact time. When the spore powder was used to fermented tempeh immediately, product has similar characters as which use spores that was fresh prepared. The tempeh has normal quality. When spore powder stored at low temperature, tempeh from storage spore in weeks 4, 8 and 12 is still normal. Time spending in production was close to the production of fresh spores. After storage spores for 16 and 20 weeks, tempeh is still normal but growth and sporulation were take longer time than usual (about 6 hours). At 24 week storage, fungal growth is not good, made tempeh looks inferior to normal color, also smell and texture.

Keywords: freez drying, preparation, spores powder, tempeh

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6 Characterization of Fungal Endophytes in Leaves, Stems and Roots of African Yam Bean (Sphenostylis sternocarpa Hochst ex. A. Rich Harms)

Authors: Iyabode A. Kehinde, Joshua O. Oyekanmi, Jumoke T. Abimbola, Olajumoke E. Ayanda

Abstract:

African yam bean (AYB), (Sphenostylis stenocarpa) is a leguminous crop that provides nutritionally rich seeds, tubers and leaves for human consumption. AYB potentials as an important food security crop is yet to be realized and thus classified as underutilized crop. Underutilization of the crop has been partly associated with scarce information on the incidence and characterization of fungal endophytes infecting vascular parts of AYB. Accurate and robust detection of these endophytic fungi is essential for diagnosis, modeling, surveillance and protection of germplasm (seed) health. This work aimed at isolating and identifying fungal endophytes associated with leaves, stems and roots of AYB in Ogun State, Nigeria. This study investigated both cultural and molecular properties of endophytic fungi in AYB for its characterization and diversity. Fungal endophytes were isolated and culturally identified. DNA extraction, PCR amplification using ITS primers and analyses of nucleotide sequences of ribosomal DNA fragments were conducted on selected isolates. BLAST analysis was conducted on consensus nucleotide sequences of 28 out of 30 isolates and results showed similar homology with genera of Rhizopus, Cunninghamella, Fusarium, Aspergillus, Penicillium, Alternaria, Diaporthe, Nigrospora, Purpureocillium, Corynespora, Magnaporthe, Macrophomina, Curvularia, Acrocalymma, Talaromyces and Simplicillium. Slight similarity was found with endophytes associated with soybean. Phylogenetic analysis by maximum likelihood method showed high diversity among the general. These organisms have high economic importance in crop improvement. For an instance, Purpureocillium lilacinum showed high potential in control of root rot caused by nematodes in tomatoes. Though some can be pathogens, but many of the fungal endophytes have beneficial attributes to plant in host health, uptake of nutrients, disease suppression, and host immunity.

Keywords: molecular characterization, African Yam Bean, fungal endophyte, plant parts

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5 Microbiological Examination and Antimicrobial Susceptibility of Microorganisms Isolated from Salt Mining Site in Ebonyi State

Authors: Anyimc, C. J. Aneke, J. O. Orji, O. Nworie, U. C. C. Egbule

Abstract:

The microbial examination and antimicrobial susceptibility profile of microorganism isolated from the salt mining site in Ebonyi state were evaluated in the present study using a standard microbiological technique. A total of 300 samples were randomly collected in three sample groups (A, B, and C) of 100 each. Isolation, Identification and characterization of organization present on the soil samples were determined by culturing, gram-staining and biochemical technique. The result showed the following organisms were isolated with their frequency as follow: Bacillus species (37.3%) and Staphylococcus species(23.5%) had the highest frequency in the whole Sample group A and B while Klebsiella specie (15.7%), Pseudomonas species(13.7%), and Erwinia species (9.8%) had the least. Rhizopus species (42.0%) and Aspergillus species (26.0%) were the highest fungi isolated, followed by Penicillum species (20.0%) while Mucor species (4.0%), and Fusarium species (8.0%) recorded the least. Sample group C showed high microbial population of all the microbial isolates when compared to sample group A and B. Disc diffusion method was used to determine the susceptibility of isolated bacteria to various antibiotics (oxfloxacin, pefloxacin, ciprorex, augumentin, gentamycin, ciproflox, septrin, ampicillin), while agar well diffusion method was used to determine the susceptibility of isolated fungi to some antifungal drugs (metronidazole, ketoconazole, itraconazole fluconazole). The antibacterial activity of the antibiotics used showed that ciproflux has the best inhibitory effect on all the test bacteria. Ketoconazole showed the highest inhibitory effect on the fungal isolates, followed by itraconazole, while metronidazole and fluconazole showed the least inhibitory effect on the entire test fungal isolates. Hence, the multiple drug resistance of most isolates to appropriate drugs of choice are of great public health concern and cells for periodic monitoring of antibiograms to detect possible changing patterns. Microbes isolated in the salt mining site can also be used as a source of gene(s) that can increase salt tolerance in different crop species through genetic engineering.

Keywords: microorganisms, antibacterial, antifungal, resistance, salt mining site, Ebonyi State

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4 Microorganisms in Fresh and Stored Bee Pollen Originated from Slovakia

Authors: Vladimíra Kňazovická, Mária Dovičičová, Miroslava Kačániová, Margita Čanigová

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The aim of the study was to test the storage of bee pollen at room temperature and in cold store, and to describe microorganisms originated from it. Fresh bee pollen originating in West Slovakia was collected in May 2010. It was tested for presence of particular microbial groups using dilution plating method, and divided into two parts with different storage (in cold store and at room temperature). Microbial analyses of pollen were repeated after one year of storage. Several bacterial strains were isolated and tested using Gram staining, for catalase and fructose-6-phosphate-phosphoketolase presence, and by rapid ID 32A (BioMérieux, France). Micromycetes were identified at genus level. Fresh pollen contained coliform bacteria, which were not detected after one year of storage in both ways. Total plate count (TPC) of aerobes and anaerobes and of yeasts in fresh bee pollen exceeded 5.00 log CFU/g. TPC of aerobes and anaerobes decreased below 2.00 log CFU/g after one year of storage in both ways. Count of yeasts decreased to 2.32 log CFU/g (at room temperature) and to 3.66 log CFU/g (in cold store). Microscopic filamentous fungi decreased from 3.41 log CFU/g (fresh bee pollen) to 1.13 log CFU/g (at room temperature) and to 1.89 log CFU/g (in cold store). In fresh bee pollen, 12 genera of micromycetes were identified in the following order according to their relative density: Penicillium > Mucor > Absidia > Cladosporium, Fusarium > Alternaria > Eurotium > Aspergillus, Rhizopus > Emericella > Arthrinium and Mycelium sterilium. After one year at room temperature, only three genera were detected in bee pollen (Penicillium > Aspergillus, Mucor) and after one year in cold store, seven genera were detected (Mucor > Penicillium, Emericella > Aspergillus, Absidia > Arthrinium, Eurotium). From the plates designated for anaerobes, eight colonies originating in fresh bee pollen were isolated. Among them, a single yeast isolate occurred. Other isolates were G+ bacteria, with a total of five rod shaped. In three out of these five, catalase was absent and fructose-6-phosphate-phosphoketolase was present. Bacterial isolates originating in fresh pollen belonged probably to genus Bifidobacterium or relative genera, but their identity was not confirmed unequivocally. In general, cold conditions are suitable for maintaining the natural properties of foodstuffs for a longer time. Slight decrease of microscopic fungal number and diversity was recorded in cold temperatures compared with storage at room temperature.

Keywords: bacteria, bee product, microscopic fungi, biosystems engineering

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3 Antimicrobial Activity of Fatty Acid Salts against Microbes for Food Safety

Authors: Aya Tanaka, Mariko Era, Manami Masuda, Yui Okuno, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

Abstract:

Objectives— Fungi and bacteria are present in a wide range of natural environments. They are breed in the foods such as vegetables and fruit, causing corruption and deterioration of these foods in some cases. Furthermore, some species of fungi and bacteria are known to cause food intoxication or allergic reactions in some individuals. To prevent fungal and bacterial contamination, various fungicides and bactericidal have been developed that inhibit fungal and bacterial growth. Fungicides and bactericides must show high antifungal and antibacterial activity, sustainable activity, and a high degree of safety. Therefore, we focused on the fatty acid salt which is the main component of soap. We focused on especially C10K and C12K. This study aimed to find the effectiveness of the fatty acid salt as antimicrobial agents for food safety. Materials and Methods— Cladosporium cladosporioides NBRC 30314, Penicillium pinophilum NBRC 6345, Aspergillus oryzae (Akita Konno store), Rhizopus oryzae NBRC 4716, Fusarium oxysporum NBRC 31631, Escherichia coli NBRC 3972, Bacillus subtilis NBRC 3335, Staphylococcus aureus NBRC 12732, Pseudomonas aenuginosa NBRC 13275 and Serratia marcescens NBRC 102204 were chosen as tested fungi and bacteria. Hartmannella vermiformis NBRC 50599 and Acanthamoeba castellanii NBRC 30010 were chosen as tested amoeba. Nine fatty acid salts including potassium caprate (C10K) and laurate (C12K) at 350 mM and pH 10.5 were used as antifungal activity. The spore suspension of each fungus (3.0×10⁴ spores/mL) or the bacterial suspension (3.0×10⁵ or 3.0×10⁶ or 3.0×10⁷ CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar or nutrient agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 °C. Results— C10K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than A. oryzae. C12K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than P. pinophilum and A. oryzae. C10K and C12K did not show high anti-yeast activity. C10K was antibacterial activity of 6 or 7 log-unit incubated time for 10 min against bacteria other than B. subtilis. C12K was antibacterial activity of 5 to 7 log-unit incubated time for 10 min against bacteria other than S. marcescens. C12K was anti-amoeba activity of 4 log-unit incubated time for 10 min against H. vermiformis. These results suggest C10K and C12K have potential in the field of food safety.

Keywords: food safety, microbes, antimicrobial, fatty acid salts

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2 Fermented Fruit and Vegetable Discard as a Source of Feeding Ingredients and Functional Additives

Authors: Jone Ibarruri, Mikel Manso, Marta Cebrián

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A high amount of food is lost or discarded in the World every year. In addition, in the last decades, an increasing demand of new alternative and sustainable sources of proteins and other valuable compounds is being observed in the food and feeding sectors and, therefore, the use of food by-products as nutrients for these purposes sounds very interesting from the environmental and economical point of view. However, the direct use of discarded fruit and vegetables that present, in general, a low protein content is not interesting as feeding ingredient except if they are used as a source of fiber for ruminants. Especially in the case of aquaculture, several alternatives to the use of fish meal and other vegetable protein sources have been extensively explored due to the scarcity of fish stocks and the unsustainability of fishing for these purposes. Fish mortality is also of great concern in this sector as this problem highly reduces their economic feasibility. So, the development of new functional and natural ingredients that could reduce the need for vaccination is also of great interest. In this work, several fermentation tests were developed at lab scale using a selected mixture of fruit and vegetable discards from a wholesale market located in the Basque Country to increase their protein content and also to produce some bioactive extracts that could be used as additives in aquaculture. Fruit and vegetable mixtures (60/40 ww) were centrifugated for humidity reduction and crushed to 2-5 mm particle size. Samples were inoculated with a selected Rhizopus oryzae strain and fermented for 7 days in controlled conditions (humidity between 65 and 75% and 28ºC) in Petri plates (120 mm) by triplicate. Obtained results indicated that the final fermented product presented a twofold protein content (from 13 to 28% d.w). Fermented product was further processed to determine their possible functionality as a feed additive. Extraction tests were carried out to obtain an ethanolic extract (60:40 ethanol: water, v.v) and remaining biomass that also could present applications in food or feed sectors. The extract presented a polyphenol content of about 27 mg GAE/gr d.w with antioxidant activity of 8.4 mg TEAC/g d.w. Remining biomass is mainly composed of fiber (51%), protein (24%) and fat (10%). Extracts also presented antibacterial activity according to the results obtained in Agar Diffusion and to the Minimum Inhibitory Concentration (MIC) tests determined against several food and fish pathogen strains. In vitro, digestibility was also assessed to obtain preliminary information about the expected effect of extraction procedure on fermented product digestibility. First results indicated that remaining biomass after extraction doesn´t seem to improve digestibility in comparison to the initial fermented product. These preliminary results show that fermented fruit and vegetables can be a useful source of functional ingredients for aquaculture applications and a substitute of other protein sources in the feeding sector. Further validation will be also carried out through “in vivo” tests with trout and bass.

Keywords: fungal solid state fermentation, protein increase, functional extracts, feed ingredients

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