Search results for: inoculated

Authors: H. M. Takematsu, B. R. De Camargo, E. F. Noronha

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The use of hydrolyzing enzymes for degradation of lignocellulosic biomass is of great concern for the production of second generation ethanol. Although many hollocelulases have already been described in the literature, much more has to be discovered. Therefore, the aim of this study to evaluate hollocelulase production of P. polonicum grown in liquid media containing sugarcane bagasse as the carbon source. From a collection of twenty fungi isolated from Cerrado biome soil, P. polonicum was molecular identified by sequencing of ITS4, βtubulin and Calmodulin genes, and has been chosen to be further investigated regarding its potential production of hydrolyzing enzymes. Spore suspension (1x10-6 ml-1) solution was inoculated in sterilized minimal liquid medium containing 0,5%(w/v) of non-pretreated sugarcane bagasse as the carbon source, and incubated in shaker incubator at 28°C and 120 rpm. The supernatant obtained, was subjected to enzymatic assays to analyze xylanase, mannanase, pectinase and endoglucanase activities. Xylanase activity showed better results (67,36 UI/mg). Xylanases bands were indicated by zymogram and SDS-PAGE, and one of them was partially purified and characterized. It showed maximum activity at 50 °C, was thermostable for 72h at 40°C, and pH5.0 was the optimum observed. This study presents P. polonicum as an interesting source of hollocelulases, especially xylanase, for lignocellulose bio-conversion processes with commercial use.

Keywords: sugarcane bagasse, Cerrado biome , hollocelulase, lignocellulosic biomass

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Authors: Ritu Chaturvedi, Manoj Paul

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A pot experiment was conducted to investigate the effect of Arbuscular mycorrhizal fungus (AMF) on metal(loid) uptake and accumulation efficiency of Pisum sativum along with physiological and biochemical response. Plants were grown in soil spiked with 50 and 100 mg kg-1 Pb, 25 and 50 mg kg-1 Cd, 50 and 100 mg kg-1 As and a combination of all three metal(loid)s. A parallel set was maintained and inoculated with arbuscular mycorrhizal fungus for comparison. After 60 days, plants were harvested and analysed for metal(loid) content. A steady increase in metal(loid) accumulation was observed on increment of metal(loid) dose and also on AMF inoculation. Plant height, biomass, chlorophyll, carotenoid and carbohydrate content reduced upon metal(loid) exposure. Increase in enzymatic (CAT, SOD and APX) and nonenzymatic (Proline) defence proteins was observed on metal(loid) exposure. AMF inoculation leads to an increase in plant height, biomass, chlorophyll, carotenoids, carbohydrate and enzymatic defence proteins (p≤0.001) under study; whereas proline content was reduced. Considering the accumulation efficiency and adaptive response of plants and alleviation of stress by AMF, this symbiosis can be applied for on-site remediation of Pb and Cd contaminated soil.

Keywords: heavy metal, mycorrhiza, pea, phyroremediation

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Authors: Mohammad Reza Haj Seyed Hadi, Fereshteh Ghanepasand, Mohammad Taghi Darzi

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The main objective of this study was to determine the effects of Nitrogen fixing bacteria and manure application on the seed yield and yield components in black cumin (Nigella sativa L.). The experiment was carried out at the RAN Research Station in Firouzkouh in 2012. A 4×4 factorial experiment, arranged in a randomized complete blocks designed with three replications. The treatments consisted of 4 level of nitrogen fixing bacteria (control, Azotobacter, Azospirillum and Azotobacter + Azospirillum) and 4 level of manure (0, 2.5, 5 and 7.5 ton ha-1). The present results have shown that the highest height, 1000 seeds weight, seed number per follicle, follicle yield, seed yield and harvest index were obtained after using Azotobacter and Azospirillum, simultaneously. Manure application only affects on follicle yield and by 5ton manure ha-1 the highest follicle yield obtained. Results of this investigation showed that the maximum seed yield obtained when Aotobacter+Azospirillum inoculated with black cumin seeds and 5 ton manure ha-1 applied. Combined application of nitrogen fixing bacteria and manure can be helpful in developing of production and yield in Black cumin.

Keywords: azotobacter, azospirillum, black cumin, yield, yield components

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Authors: F. Motamedi Sedeh, Sh. Chahardoli, H. Mahravani, N. Harzandi, M. Sotoodeh, S. K. Shafaei

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Foot-and-mouth disease virus (FMDV) is the most economically important disease of livestock. The aim of the study is inactivation of FMD virus type O/IRN/2010 by electron beam without antigenic changes as electron radio vaccine. The BALB/C mice were divided into three groups, each group containing five mice. Three groups of mice were inoculated with conventional vaccine and electron beam irradiated vaccine FMDV type O/IRN/2010 subcutaneously three weeks interval, the final group as negative control. The sera were separated from the blood samples of mice 14 days after last vaccination and tested for the presence of antibodies against FMDV type O/IRN/2010 by serum neutralization test. The Serum Neutralization Test (SNT) was carried out and antibody titration was calculated according to the Kraber protocol. The results of the SNT in three groups of mice showed the titration of neutralizing antibody in the vaccinated mice groups; electron radio vaccine and conventional vaccine were significantly higher than negative control group (P<0.05). Therefore, the radio vaccine is a good candidate to immunize animals against FMDV type O/IRN/2010.

Keywords: FMDV type O/IRN/2010, neutralizing antibody response, electron beam, radio vaccine

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Authors: Cigdem Yangin-Gomec, Aigerim Jaxybayeva, Orhan Ince

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In this study, the anaerobic treatability of chicken manure diluted with tap water (with an influent feed ratio of 1 kg of fresh chicken manure to 6 liter of tap water) was investigated in a lab-scale anaerobic sludge bed (ASB) reactor inoculated with the granular sludge already adapted to chicken manure. The raw waste digested in this study was the manure from laying-hens having average total solids (TS) of about 30% with ca. 60% volatile content. The ASB reactor was fed semi-continuously at ambient operating temperature range (17-23^◦C) at a HRT of 13 and 26 days for about 6 months, respectively. The respective average total and soluble chemical oxygen demand (COD) removals were ca. 90% and 75%, whereas average biomethane production rate was calculated ca. 180 lt per kg of COD_removed from the ASB reactor at an average HRT of 13 days. Moreover, total suspended solids (TSS) and volatile suspended solids (VSS) in the influent were reduced more than 97%. Hence, high removals of the organic compounds with respective biogas production made anaerobic stabilization of the diluted chicken manure by ASB reactor at ambient operating temperatures viable. By this way, external heating up to 35^◦C (i.e. anaerobic processes have been traditionally operated at mesophilic conditions) could be avoided in the scope of this study.

Keywords: ambient anaerobic digestion, biogas recovery, poultry manure, renewable energy

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Authors: F. Haddadj, F. Acheuk, S. Hamdi, S. Zenia, A. Smai, H. Saadi, B. Doumandji-Mitiche

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Locust is causing significant losses in agricultural production in the countries concerned by the invasion. Up to the present control strategy has consisted only of the spreaders chemicals; they have proven harmful to the environment and. For this, a new control method appeared it comes to the biological control based mostly by using microorganism. It is in that sense is we've made our contribution by the use of a biopesticide which is entomopathogenic fungus Metarhizium anisopliae var acridium "Green Muscle" on part of the cuticule the larval of fifth instar locust Schistocerca gregaria (Forskål, 1775). Preliminary test on the study of the pathogenicity of M. anisopliae var acridium biocontrol agent, was conducted in the laboratory on L5 S. gregaria, on which we inoculated treatment in the digestive tract and it administrant 20μl of entomopathogenic solution orally at a dose DL50 = 3.25 x107 sp./ ml (median lethal dose estimated at earlier), 5 days after treatment individuals are sacrificed. After dissection cuticles are recovered and then subjected to histological sections. The histological technique followed is that of Martoja Martoja-Pierson (1967). Microscopic observation revealed alterations in the architecture of the cuticule which leads to disorganization of cell layers.

Keywords: biopesticide, cuticle, desert locust, toxicity

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Authors: Yousry El-Sayed Elbolkiny, Mohamed Labib Salem

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Background: Rosemary (ROLE) and costus (SLRE) have been established to show antioxidant effects. Aim: This study aimed to evaluate the ameliorating effects of ROLE and SLRE on the side effects induced by cisplatin (CIS) in tumor-bearing mice. Materials and Methods: Extracts of ROLE and SLRE were examined for their phytochemical activities. To evaluate their anti-tumor effects, mice were inoculated intraperitoneally (i.p.) with 2.5x105 Ehrlich ascites carcinoma (EAC) and then treated i.p. with CIS at days 3-7 and with ROLE (dose) or SLRE (dose) at days 3-14. Mice were sacrificed on day 14 for CBC and T-cell analyses. Results: Phytochemical analysis revealed that both ROLE and SLRE showed similar antioxidant activities. Treatment of EAC-bearing mice with CIS-induced antitumor efficacy of about 90%. Treatment with CIS in combination with ROLE or SLRE did not further enhance the antitumor activity of CIS. However, co-administration of ROLE or SLRE with CIS significantly increased the antitumor efficacy of CIS. Flow cytometric analysis showed that the numbers of CD4+ and CD8+ T cells were decreased in EAC-bearing mice after treatment with CIS. Treatment with both ROLE and SLRE improved the number of these cells. Conclusion: Combinatorial treatment with rosemary and costus can enhance the antitumor activity of CIS

Keywords: CBC, cisplantin, costus, rosemary

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Authors: Sang Guen Kim, Sib Sankar Giri, Jin Woo Jun, Saekil Yun, Hyoun Joong Kim, Sang Wha Kim, Jung Woo Kang, Se Jin Han, Se Chang Park

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Emerging of the super bacteria, bacteriophages are considered to be as an alternative to antibiotics. As the demand of phage increased, economical and large production of phage is becoming one of the critical points. For the therapeutic use, what is important is to eradicate the pathogenic bacteria as fast as possible, so higher concentration of phages is generally needed for effective therapeutic function. On the contrary, for the maximum production, bacteria work as a phage producing factory. As a microbial cell factory, bacteria is needed to last longer producing the phages without eradication. Consequently, killing the bacteria fast has a negative effect on large production. In this study, Multiplicity of Infection (MOI) was manipulated based on initial bacterial inoculation and used phage pAh-6C which has therapeutic effect against Aeromonas hydrophila. 1, 5 and 10 percent of overnight bacterial culture was inoculated and each bacterial culture was co-cultured with the phage of which MOI of 0.01, 0.0001, and 0.000001 respectively. Simply changing the initial MOI as well as bacterial inoculation concentration has regulated the production quantity of the phage without any other changes to culture conditions. It is anticipated that this result can be used as a foundational data for mass production of lytic bacteriophages which can be used as the therapeutic bio-control agent.

Keywords: bacteriophage, multiplicity of infection, optimization, Aeromonas hydrophila

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Authors: U.N. Emiri, E. Moroyei

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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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Authors: Nawel Boucherba, Azzedine Bettache, Abdelaziz Messis, Francis Duchiron, Said Benallaoua

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The impetus for research in the field of bioseparation has been sparked by the difficulty and complexity in the downstream processing of biological products. Indeed, 50% to 90% of the production cost for a typical biological product resides in the purification strategy. There is a need for efficient and economical large scale bioseparation techniques which will achieve high purity and high recovery while maintaining the biological activity of the molecule. One such purification technique which meets these criteria involves the partitioning of biomolecules between two immiscible phases in an aqueous system (ATPS). The Production of xylanases is carried out in 500ml of a liquid medium based on birchwood xylan. In each ATPS, PEG 1000 is added to a mixture consisting of dipotassium phosphate, sodium chloride and the culture medium inoculated with the strain Jonesia denitrificans, the mixture was adjusted to different pH. The concentration of PEG 1000 was varied: 8 to 16 % and the NaCl percentages are also varied from 2 to 4% while maintaining the other parameters constant. The results showed that the best ATPS for purification of xylanases is composed of PEG 1000 at 8.33%, 13.14 % of K2HPO4, 1.62% NaCl at pH 7. We obtained a yield of 96.62 %, a partition coefficient of 86.66 and a purification factor of 2.9. The zymogram showed that the activity is mainly detected in the top phase.

Keywords: Jonesia denitrificans BN13, xylanase, aqueous two phases system, zymogram

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Authors: Isa Shu’aibu, A. A. Mu’inat, F. U. Maigari, M. A. Mani

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Candida albicans is the common cause of both oral and vaginal candidiasis in humans. This candidiasis leads to a wide range of physical, psychological and even physiological problems in humans particularly pregnant women. Samples of endocervical and high vaginal swab were collected from 200 women attending Gombe Specialist Hospital and inoculated on Saboraud Dextrose Agar (SDA) incorporated with chloramphenicol to get rid of the unwanted bacterial contaminants. Gram staining technique and germ tube test were employed for the identification, as Candida albicans is positive for both. Gram positive samples were 70% (n=140) and were further subjected to germ tube test. The remaining 30% (n=60) were found to be Gram negative. 90% (n=126) of the Gram positive ones isolated were also found to be positive for germ tube test; confirming the presence of Candida albicans. Antifungal susceptibility testing revealed that members of Imidazole (Ketoconazole, Miconazole) and those of Triazoles (Fluconazole and Itraconazole) were found to be more effective at concentrations of 20, 50 and 100 µg/disc compared to Griseofulvin (Fulcin) with only 26.00 mm zone of inhibition at 100 µg/disc concentration.

Keywords: Candida albicans, candidiasis, endocervical, vaginal swab, antifungal susceptibility, imidazole, triazoles

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Authors: Kehinde T. Kareem

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The need to increase the production of cucumber has led to the use of inorganic fertilizers. This chemical affects the ecological balance of nature by increasing the nitrogen and phosphorus contents of the soil. Surface runoffs into rivers and streams cause eutrophication which affects aquatic organisms as well as the consumers of aquatic animals. Therefore, this study was carried out in the screenhouse to investigate the use of a plant growth-promoting fungus; Trichoderma longibrachiatum for the growth promotion of conventional and in-vitro propagated Ashley and Marketmoor cucumber. Before planting of cucumber, spore suspension (108 cfu/ml) of Trichoderma longibrachiatum grown on Potato dextrose agar (PDA) was inoculated into the soil. Fruits were evaluated for the presence of Trichoderma longibrachiatum using a species-specific primer. Results revealed that the highest significant plant height produced by in-vitro propagated Ashley was 19 cm while the highest plant height of in-vitro propagated Marketmoor was 19.67 cm. The yield of the conventional propagated Ashley cucumber showed that the number of fruit/plant obtained from T. longibrachiatum-fertilized plants were significantly more than those of the control. The in-vitro Ashely had 7 fruits/plant while the control produced 4 fruits/plant. In-vitro Marketmoor had ten fruits/plant, and the control had a value of 4 fruits/plant. There were no traces of Trichoderma longibrachiatum genes in the harvested cucumber fruits. Therefore, the use of Trichoderma longibrachiatum as a plant growth-promoter is safe for human health as well as the environment.

Keywords: biofertilizer, cucumber, genes, growth-promoter, in-vitro, propagation

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Authors: Vinicius Buccelli Ribeiro, Maria Teresa Destro, Mariza Landgraf

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Bacteriophages are one of the most abundant replicating entities on Earth and can be found everywhere in which their hosts live and there are reports regarding isolation from different niches such as soil and foods. Since studies are moving forward with regard to biotechnology area, different research projects are being performed focusing on the phage technology and its use by the food industry. This study aimed to evaluate a cocktail (LP501) of phages isolated in Brazil for its lytic potential against Listeria monocytogenes. Three bacteriophages (LP05, LP12 and LP20) were isolated from soil samples and all of them showed 100% lysis against a panel of 10 L. monocytogenes strains representing different serotypes of this pathogen. A mix of L. monocytogenes 1/2a and 4b were inoculated in soft cheeses (approximately 105 cfu/cm2) with the phage cocktail added thereafter (1 x 109 PFU/cm2). Samples were analyzed immediately and then stored at 10°C for ten days. At 30 min post-infection, the cocktail reduced L. monocytogenes counts approximately 1.5 logs, compared to controls without bacteriophage. The treatment produced a statistically significant decrease in the counts of viable cells (p < 0.05) and in all assays performed we observed a decrease of up to 4 logs of L. monocytogenes. This study will make available to the international community behavioral and molecular data regarding bacteriophages present in soil samples in Brazil. Furthermore, there is the possibility to apply this new cocktail of phages in different food products to combat L. monocytogenes.

Keywords: bacteriophages, biocontrol, listeria monocytogenes, soft cheese

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Authors: Ezeonu, Chukwuma Stephen, Onwurah, Ikechukwu Noel Emmanuel

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Pure strains of Aspergillus fumigatus (AF), aspergillus niger (AN), aspergillus oryzae (AO), trichophyton mentagrophyte (TM), trichophyton rubrum (TR) and Trichophyton soudanense (TS) were isolated from decomposing rice husk. Freshly processed rice husk in Mandle’s medium were heat pre-treated using an autoclave at 121oC for 20 minutes. The isolated fungi as monoculture and di-culture combinations were inoculated into each of the pre-treated rice husk with the exception of two controls. Seven days hydrolysis was followed by estimation of carbohydrate, reducing sugar and non-reducing sugar. Fungal treated rice husks were left to ferment for 7 days with introduction of both baker’s and palm wine yeast. The result obtained in the work gave the highest carbohydrate (20.53 ± 2.73 %) from rice husks treated with TS + TR di-culture. The highest soluble reducing sugar (2.66 ± 0.14 %) was obtained from rice husk treated with TM. The highest soluble nonreducing sugar (18.08 ± 2.61 %) was from AF. The introduction of yeasts from palm wine gave the highest bio-ethanol (12.82 ± 0.39 %) from AO. The highest bio-ethanol (6.60 ± 0.10 %) from baker's yeast fermentation was in AO + TS treated rice husk. There was increased availability of sugar and moderate yield of bio-ethanol, especially from palm wine yeast.

Keywords: fungi, rice husk, carbohydrate, reducing sugar, non-reducing sugar, ethanol, fermentation

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Authors: Mostafa A. Amer, I. A. El-Samra, I. I. Abou-ElSeoud, S. M. El-Abd, N. K. Shawertamimi

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Tomato Fusarium wilt disease caused by Fusarium oxysporum f. sp. Lycopercisi (FOL) is considered one of the most destructive diseases in Egypt. Effect of some biotic inducers such as Bacillus megaterium var. phosphaticum, Glomus intraradices and Glomus macrocarpum at seven different mixed treatments, was tested for their ability to induce resistance in tomato plants against the disease. According to pathogenicity tests, all the tested isolates of FOL showed wilt symptoms on both of the tested cultivars; however, they considerably varied in percentages of disease incidence (DI) and disease severity (DS). Castle Rock was more susceptible than Peto 86, which was relatively resistant. Pretreatment of both cultivars, under greenhouse conditions, with the tested biotic inducers alone or in combination with each other's, significantly increased the induction of chitinase, β-1,3-glucanase, peroxidase, and polyphenoloxidase and reduced disease incidence and severity, compared with untreated noninoculated (C1) and untreated inoculated (C2) controls. Application of a combination of BMP, with GI and GM was the most effective in increasing the induction rated of the tested enzymes, compared with the other treatments. Induction of enzymes in most of the tested bioinducers treatments gradually increased, attaining maximum values after 48 or/and 72 hrs after challenging with FOL, then gradually declined. GI was the least effective bioinducer.

Keywords: F. oxysporum f. sp. lycopersici, defense enzymes, induced systemic resistance, ISR, B. megaterium var. phosphaticum, G. macrocarpum, G. intraradices

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Authors: Sadhana Shukla, Pushplata Singh, Nidhi Didwania

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Arbuscular mycorrhizal (AM) fungi are a highly advantageous and versatile group of fungi that significantly contribute to the formation of soil organic matter by creating a demand for plant carbon (C) and distributing it through below-ground hyphal biomass, regardless of their substantial contribution in enhancing net primary productivity and accumulating additional photosynthetic fixed C in the soil. The genetic role of AM fungi in carbon cycling is largely unexplored. In our study, we propose that AM fungi significantly interact with the soil, particularly: the provision of photosynthates by plants. We have studied the expression of AM fungi genes involved in CO₂ sequestration during host-plant interaction was investigated by qPCR studies. We selected Rhizophagus proliferus (AM fungi) and Oryza sativa (Rice) (inoculated with or without 200ppg AMF inoculums per plant) and investigated the effect of AM fungi on soil organic carbon (SOC) and rice growth under field conditions. Results thus provided faster SOC turnover, 35% increased nutrient uptake in plants and pronounced hyphal biomass of AM fungi which enhanced soil carbon storage by 15% in comparison to uninoculated plants. This study will offer a foundation for delving into various carbon-soil studies while also advancing our comprehension of the relationship between AM fungi and the sustainability of agricultural ecosystems.

Keywords: arbuscular mycorrhizal (AM) fungi, carbon sequestration, gene expression, soil health, plant development.

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Authors: Ahmad Alazemi, M. Sherif El-Eskandrany, Mohamad Kishk, Thanyan AlOnaizi, Ahmad Alduweesh, Shorouq Abdullaleel

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Arc melting technique followed by top-down approach, using a high-energy ball milling technique were employed to synthesize nanocrystalline of Cu50(Zr50-xNix)50 (x = 0, 10, 20 and 30 at.%) powder particles. The end-products of the alloy powders obtained after 50 h of the ball milling time were uniform in composition and had spherical-like morphology with an average particle size of 0.75 µm in diameter. The powders, which consisted of nanocrystalline grains with an average grain size of 10 nm in diameter, were used as feedstock materials for double face coating of stainless (SUS304) sheets, using cold spraying process. The coating materials enjoyed nanocrystalline structure and uniform composition. Biofilms were grown on 20-mm2 SUS304 sheets coated coupons inoculated with 1.5 × 108 CFU ml−1 E. coli. Significant biofilm inhibition was recorded in the nanoparticles coated coupons in comparison to non-coated SUS304 coupon. In conclusion, this study demonstrates that formation of biofilms can be significantly inhibited by Cu-based alloys especially in case of high (Ni) content. The inhibition of biofilm formation by nanocrystalline powders of Cu-based provides a practical approach to achieve the inhibition of biofilms formed by an emerging pathogen.

Keywords: biofilm, Cu, E.coli, FE-HRTEM/EDS, nanomaterials, nanocrystalline

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Authors: Mohammad Hajjartabar, Tahereh Kermani Ranjbar

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P. aeruginosa EF2 was isolated and identified from human infection sources before in our previous study. The present study was performed to determine the characteristics and activity role of bioflocculant produced by the bacterium in flocculation of the wastewater active sludge treatment. The bacterium was inoculated and then was grown in an orbital shaker at 250 rpm for 5 days at 35 °C under TSB and peptone water media. After incubation period, culture broths of the bacterial strain was collected and washed. The concentration of the bacteria was adjusted. For the extraction of the bacterial bioflocculant, culture was centrifuged at 6000 rpm for 20 min at 4 °C to remove bacterial cells. Supernatant was decanted and pellet containing bioflocculant was dried at 105 °C to a constant weight according to APHA, 2005. The chemical composition of the extracted bioflocculant from the bacterial sample was then analyzed. Wastewater active sludge sample obtained from aeration tank from one of wastewater treatment plants in Tehran, was first mixed thoroughly. After addition of bioflocculant, improvements in floc density were observed with an increase in bioflocculant. The results of this study strongly suggested that the extracted bioflucculant played a significant role in flocculation of the wastewater sample. The use of wild bacteria and nutrient regulation techniques instead of genetic manipulation opens wide investigation area in the future to improve wastewater treatment processes. Also this may put a new path in front of us to attain and improve the more effective bioflocculant using the purified microbial culture in wastewater treatment.

Keywords: wastewater treatment, P. aeruginosa, sludge treatment

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Authors: S. Mantzoukis, M. Gerasimou, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

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Purpose: Patients in Intensive Care Units (ICU) are exposed to a different spectrum of microorganisms relative to the hospital. Due to the fact that the majority of these patients are intubated, bronchial secretions should be examined. Material and Method: Bronchial secretions should be taken with care so as not to be mixed with sputum or saliva. The bronchial secretions are placed in a sterile container and then inoculated into blood, Mac Conkey No2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. After this period, if any number of microbial colonies are detected, gram staining is performed and then the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) followed by a sensitivity test in the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017 the Laboratory of Microbiology received 365 samples of bronchial secretions from the Intensive Care Unit. 237 were found positive. S. epidermidis was identified in 1 specimen, A. baumannii in 60, K. pneumoniae in 42, P. aeruginosa in 50, C. albicans in 40, P. mirabilis in 4, E. coli in 4, S. maltophilia in 6, S. marcescens in 6, S. aureus in 12, S. pneumoniae in 1, S. haemolyticus in 4, P. fluorescens in 1, E. aerogenes in 1, E. cloacae in 5. Conclusions: The majority of ICU patients appear to be a fertile ground for the development of infections. The nature of the findings suggests that a significant part of the bacteria found comes from the unit (nosocomial infection).

Keywords: bronchial secretions, cultures, infections, intensive care units

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Authors: S. S. R. Shojaei, S. Kord Afshari

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The study was initiated to compare the production and defecation of Eimerial oocysts of two internationally reputed broiler strains under the local environmental and management conditions. 40 one-day old male chickens of Arbor Acres strain and ROSS strain (20 chicks from each strain) used in this study and were divided randomly into four control and challenge groups. Feed and water were provided for ad libitum consumption. At 15 d of age, chickens of challenge groups (from each strain) were individually inoculated with a mixture of 50000 of sporulated oocysts of 4 species including of E. acervulina (20%), E. maxima (40%), E. tenella (25%) and E. necatrix (15%) and also from the fourth day after Eimerial challenge, faecal droppings (litter samples) were collected 10 days consecutively for counting oocyst per gram (OPG). The results indicated that in the challenge groups, there was an increasing process of OPG in days of 4 to 7 post challenging and the pick level of OPG was seen at seventh day after challenging. From day 8 to 9, decreasing of OPG was happened. This decreasing continues with mild, fast and mild process to day of 13. Respectively and totally the average of OPG in the Arbor Acres group was lower than the group Ross in all days post inoculation and this difference was significant according to t-test. According to the obtained results in this study and since oocyst index almost always is considered as one of the most important indicators for coccidiosis evaluation, it can be realized that in the same surveillance condition the regarding the severity evaluation of coccidiosis, Arbor Acres strain broilers shed less oocysts than Ross strain broilers.

Keywords: arbor acres, ross, coccidiosis, OPG

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Authors: Djoudi Abdelhak, Djibaoui Rachid, Reguieg Yassaad Houcine

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Our study focuses on the identification of some species of Pseudomonas (P4, P5, P7 and P8) isolated from saline soils in northwestern Algeria and the effect of their metabolites on the growth of Alternaria alternata the causative agent of the blight of the bean disease (Vicia faba). We are also interested in stimulating the growth of this plant species in saline conditions (60 mM/l NaCl) and the absence of salts. The analysis focuses on rates of inhibition of mycelial growth of Alternaria alternata strain and the rate of growth of plants inoculated with strains of Pseudomonas expressed by biometrics. According to the results of the in-vitro test, P5 and P8 species and their metabolites showed a significant effect on mycelia growth and production of spores of Alternaria alternata. The in-vivo test shows that the species P8 and P5 were significantly and positively influencing the growth in biometric parameters of the bean in saline and salt-free condition. Inoculation with strain P5 has promoted the growth of the bean in stem height, stem fresh weight and dry weight of stems of 108.59%, 115.28%, 104.33%, respectively, in the presence of salt Inoculation with strain P5 has fostered the growth of the bean stem fresh weight of 112.47% in the presence of salt The effect of Pseudomonas species on the development of Vicia faba and the growth of Alternaria alternata is considering new techniques and methods of biological production and crop protection.

Keywords: pseudomonas, vicia faba, alternaria alternata, promoting of plant growth

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Authors: Yasaman Sanayei, Alireza Bahiraie

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This paper presents a systematic methodology based on the application of artificial neural networks for sequencing batch reactor (SBR). The SBR is a fill-and-draw biological wastewater technology, which is specially suited for nutrient removal. Employing reactive dye by Sphingomonas paucimobilis bacteria at sequence batch reactor is a novel approach of dye removal. The influent COD, MLVSS, and reaction time were selected as the process inputs and the effluent COD and BOD as the process outputs. The best possible result for the discrete pole parameter was a= 0.44. In orderto adjust the parameters of ANN, the Levenberg-Marquardt (LM) algorithm was employed. The results predicted by the model were compared to the experimental data and showed a high correlation with R2> 0.99 and a low mean absolute error (MAE). The results from this study reveal that the developed model is accurate and efficacious in predicting COD and BOD parameters of the dye-containing wastewater treated by SBR. The proposed modeling approach can be applied to other industrial wastewater treatment systems to predict effluent characteristics. Note that SBR are normally operated with constant predefined duration of the stages, thus, resulting in low efficient operation. Data obtained from the on-line electronic sensors installed in the SBR and from the control quality laboratory analysis have been used to develop the optimal architecture of two different ANN. The results have shown that the developed models can be used as efficient and cost-effective predictive tools for the system analysed.

Keywords: artificial neural network, COD removal, SBR, Sphingomonas paucimobilis

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Authors: Adria Rae Abigail R. Eda, Arvin C. Diesmos, Vance T. Vredenburg, Merab A. Chan

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Mitigating strategies using symbiotic cutaneous bacteria is one of the major concerns in the conservation of amphibian population. Batrachochytrium dendrobatidis is the causative agent of chytridiomycosis associated with mass mortality and amphibian extinctions worldwide. In the Philippines, there is a lack of study on the cutaneous bacteria of Philippine amphibians that may have beneficial effects to ward off the deadly fungal infection. In this study, cutaneous bacteria from frogs were isolated and examined for anti-B. dendrobatidis activity. Eight species of frogs were collected at Mt. Palay-palay Mataas na Gulod National Park in Cavite, a site positive for the presence of B. dendrobatidis. Bacteria were isolated from the skin of frogs by swabbing the surfaces of the body and inoculated in Reasoner´s 2A (R2A) agar. Isolated bacteria were tested for potential inhibitory properties against B. dendrobatidis through zoospore inhibition assay. Results showed that frog cutaneous bacteria significantly inhibited the growth of B. dendrobatidis in vitro. By means of 16S rRNA gene primers, the anti-B. dendrobatidis bacteria were identified to be Enterobacter sp., Alcaligenes faecalis and Pseudomonas sp. Cutaneous bacteria namely Enterobacter sp. (isolates PLd33 and PCv4) and Pseudomonas (isolate PLd31) remarkably cleared the growth of B. dendrobatidis zoospore in 1% tryptone agar. Therefore, frog cutaneous bacteria inhibited B. dendrobatidis in vitro and could possibly contribute to the immunity and defense of frogs against the lethal chytridiomycosis.

Keywords: Batrachochytrium dendrobatidis, cutaneous bacteria, frogs, zoospore inhibition assay

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Authors: Supawadee Phetkhajone, Wisuwat Songnuan

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Metalaxyl is one of the most common and effective fungicides used to control Phytophthora palmivora infection in durian (Durio zibethinus L.). The efficacy of metalaxyl residue in durian under greenhouse condition was evaluated using bioassay. Durian seedlings were treated with 2 methods of application, spraying, and soil drenching of metalaxyl, at recommended concentration (1000 mg/L). Mock treated samples were treated with 0.1% Tween20 and water for spraying and soil drenching methods, respectively. The experiment was performed in triplicates. Leaves were detached from treated plants at 0, 1, 7, 15, 20, 30, and 60 days after application, inoculated with metalaxyl-resistant and metalaxyl-sensitive isolates of P. palmivora, and incubated in a high humidity chamber for 5 days at room temperature. Metalaxyl efficacy was determined by measuring the lesion size on metalaxyl treated and mock treated samples. The results showed that metalaxyl can control metalaxyl-sensitive isolate of P. palmivora for at least 30 days after application in both methods of application. The metalaxyl-resistant isolate was not inhibited in all treatments. Leaf samples from spraying method showed larger lesions compared to soil drench method. These results demonstrated that metalaxyl applications, especially soil drenching methods showed high efficacy to control metalaxyl-sensitive isolates of P. palmivora, although it cannot control metalaxyl-resistant isolates of P. palmivora in all treatments. These qualitative data indicate that metalaxyl may suitable to control metalaxyl-sensitive isolates of P. palmivora infection.

Keywords: bioassay, degradation, durian, metalaxyl

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Authors: Fuad Ameen, Ali A. Al-Homaidan

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Biochar can be used in numerous biotechnological applications due to its properties to adsorb beneficial nutrients and harmful pollutants. Objectives: We aimed to treat heavy metal polluted organic wastes using vermicomposting process and produce a fertilizer that can be used in agriculture. We improved the process by adding biochar as well as microbial inoculum and biomass into household waste or sewage sludge before vermicomposting. The earthworm Eisenia fetida used in vermicomposting was included to accumulate heavy metals, biochar to adsorb heavy metals, and the microalga Navicula sp. or the mangrove fungus Acrophialophora sp. to promote plant growth in the final product used as a seedbed for Solanaceae vegetables. We carried out vermicomposting treatments to see the effect of different amendments. Final compost quality was analyzed for maturity. The earthworms were studied for their vitality, heavy metal accumulation, and metallothionein protein content to verify their role in the process. The compost was used as a seedbed for vegetables that were inoculated with a phytopathogen Pythium sp. known to cause root rot and destroy seeds. Compost as seedbed promoted plant growth and reduced disease symptoms in leaves. In the treatment where E. fetida, 6% biochar, and Navicula sp. had been added, 90% of the seeds germinated, while less than 20% germinated in the control treatment. The experimental plants had acquired resistance against Pythium sp. The metagenomic profile of microbial communities will be reported.

Keywords: organic wastes, vermicomposting process, biochar, mangrove fungus

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Authors: Akbar Esmaeili, Zahra Salarieh

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Cerium oxide nanoparticles (nano-series) are used as catalysts in industrial applications due to their free radical scavenging properties. Given that free radicals play an essential role in the pathology of many neurological diseases, we investigated the use of nanocrystals as a potential therapeutic agent for oxidative damage. This project synthesized nano-series from a new and environmentally friendly bio-pathway. Investigation of cerium nitrate in culture medium containing inoculated Lactobacillus acidophilus strain before incubation produces nano-series. Loaded with glatiramer acetate (GA) was formed by coating carboxymethylcellulose (CMC) and CeO2. FE-SEM analysis showed nano-series in the 9-11 nm range, spherical shape, and uniform particle size distribution. Cubic nanoparticles containing anti-multiple sclerosis (anti-Ms) treatment called GA were used. Glycerol monostearate (GMS) was used as a fat base, and evening primrose extract was used as an anti-inflammatory in cubosomes. Design-Expert® software was used to study the effects of different formulation factors on the properties of GAloaded cubic dispersions. Thirty GA-labeled cubic dispersions were prepared with GA-labeled carboxymethylcellulose and evaluated in vitro. The results showed an average nano-series size of 89.02 and a zeta potential of -49.9. Cubosomes containing GA-CMC/CeO2 showed a stable release profile for 180 min. The results showed that cubosomes containing GA-CMC/CeO2 could be a promising drug carrier with normal release behavior.

Keywords: ciochemistry, biotechnology, molecular, biology

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Authors: Richard Nyanzi, Maupi E. Letsoalo, Jacobus N. Eloff, Piet J. Jooste

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Candida albicans naturally occurs in the gastrointestinal tract (GIT) of more than 50% of humans. Overgrowth of the fungus causes several forms of candidiasis including oral thrush. Overgrowth tends to occur in immunocompromised humans such as diabetic, cancer and HIV patients. Antifungal treatment is available, but not without shortcomings. In this study, inhibitory activity of five probiotic Lactobacillus strains was demonstrated against the growth of seven clinical strains of Candida albicans by co-culturing of the organisms in a maize gruel (MG) medium. Phenotypic tests, molecular techniques and phylogenetic analysis have enabled precise identification of the organisms used in the study. The quantitative pour plate technique was used to enumerate colonies of the yeasts and the lactobacilli and the Kruskal-Wallis test and ANOVA tests were employed to compare the distributions of the colonies of the organisms. The cereal medium, containing added carbon sources, was inoculated with a Candida and a Lactobacillus strain in combination and incubated at 37 °C for 168 h. Aliquots were regularly taken and subjected to pH determination and colony enumeration. Certain Lactobacillus strains proved to be inhibitory and also lethal to some Candida albicans strains. A low pH due to Lactobacillus acid production resulted in significant low Candida colony counts. Higher Lactobacillus colony counts did not necessarily result in lower Candida counts suggesting that inhibitory factors besides low pH and competitive growth by lactobacilli contributed to the reduction in Candida counts. Such anti-Candida efficacy however needs to be confirmed by in vivo studies.

Keywords: candida albicans, oral thrush, candidiasis, lactobacillus, probiotics

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Authors: Phillip Minnaar, Neil Jolly, Louisa Beukes, Santiago Benito-Saez

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Dovyalis caffra is a tree found on the African continent. Limited information exists on the effect of acetous fermentation on the phytochemicals of Kei-apple fruit. The phytochemical content of vinegars is derived from compounds present in the fruit the vinegar is made of. Kei-apple fruit juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation (AF). Acetous fermentation followed AF, using an acetic acid bacteria consortium as an inoculant. Juice had the lowest pH and highest total acidity (TA). The wine had the highest pH and vinegars lowest TA. Total soluble solids and L-malic acid decreased during AF and acetous fermentation. Volatile acidity concentration was not different among vinegars. Gallic, syringic, caffeic, p-coumaric, and chlorogenic acids increased during acetous fermentation, whereas ferulic, sinapic, and protocatechuic acids decreased. Chlorogenic acid was the most abundant phenolic acid in both wines and vinegars. It is evident from this investigation that Kei-apple vinegar is a source of plant-derived phenolics, which evolved through fermentation. However, the AAB selection showed minimal performance with respect to VA production. Acetic acid bacteria selection for acetous fermentation should be reconsidered, and the reasons for the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, acetous fermentation, liquid chromatography, phenolic acids

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Authors: Sarunpron Khruengsai, Patcharee Pripdeevech

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This study aimed to investigate the fumigation activities of the volatile compounds produced by Trichoderma spp. against Fusarium oxysporum and F. proliferatum fungi that cause significant rot in fresh chilies. Two Trichoderma spp. were isolated from the leaves of Schefflera leucantha grown in Thailand and later identified as T. afroharzianum MFLUCC19-0090 and T. afroharzianum MFLUCC19-0091. Both in vitro and in vivo dual culture volatile assays were used to study the effects of the produced volatile compounds on mycelial growth. In vitro results showed that the volatile compounds produced by T. afroharzianum MFLUCC19-0090 significantly inhibited the growth of F. oxysporum, while the volatile compounds produced by T. afroharzianum MFLUCC19-0091 significantly inhibited the growth of F. proliferatum. The effectiveness of Trichoderma-derived volatile compounds in inhibiting the mycelial growth of the selected pathogens in the inoculated, fresh chili samples was further demonstrated in vivo. The volatile profiles of both Trichoderma spp. were characterized using gas chromatography-mass spectrometry. Seventy-three volatile compounds were detected from both strains. Among the major volatile compounds detected, phenyl ethyl alcohol was found to possess the strongest antifungal activity against both pathogens. The results support the possibility of using volatile compounds produced by T. afroharzianum MFLUCC19-0090 and T. afroharzianum MFLUCC19-0091 as alternative fumigants for preventing Fusarium rot of fresh chilies during the post-harvest period.

Keywords: antifungal activity, biocontrol, endophytic fungi, post-harvest

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Authors: Siti Helmyati, Euis Nurdiyawati, Joko Susilo, Endri Yuliati, Siti Fadhilatun Nashriyah, Kurnia Widyastuti

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Background: Iron fortification is one of the most effective and sustainable strategies to overcome anemia. It contradictively, has negative effect on gut microbiota balance. Pathogenic bacteria required iron for their growth. The iron source have greatly affect iron absorption in the intestine. Probiotic can inhibit the growth of pathogen. Lactobacillus plantarum Dad 13, Indonesian local isolate provides many benefits for health while fructo-oligosaccharides (FOS) provides selective substrates for probiotics’ growth. Objective: To determine the effect of iron fortification (NaFeEDTA and FeSO4) on antibacterial activity of synbiotic fermented milk. Methods: The antibacterial activity test was performed using the disc diffusion method. Paper discs were soaked in three kinds of synbiotic fermented milk, which are: 1) fortified with NaFeEDTA, 2) FeSO4 and 3) control. Escherichia coli was inoculated on nutrient agar medium. The ability of inhibition was shown by the formation of clear zone around the paper disc and measured in diameter (mm). Results: Synbiotic fermented milk fortified with iron (either NaFeEDTA or FeSO4) had antibacterial activity against Escherichia coli with diameter of clear zone were 6.53 mm and 12.3 mm, respectively (p<0.05). Compared to control (10.73 mm), synbiotic fermented milk fortified with FeSO4 had similar antibacterial activity (p>0.05). Conclusions: In vitro, synbiotic fermented milk fortified with NaFeEDTA and FeSO4 had different antibacterial activity against Escherichia coli. Iron fortification compound affected the antibacterial activity of synbiotic fermented milk.

Keywords: lactobacillus plantarum Dad 13, FOS, NaFeEDTA, FeSO4, antibacterial activity

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