Search results for: Bacillus cereus strain JMG-01

Authors: Ifueko Oghogho Ukponmwan, Mike O. Orji

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This study investigated the antifungal activity of Bacilluss subtilis in the control of postharvest fungal rot of white yam (Dioscorea spp). Bacillus subtilis was isolated from the soil and fungi (Aspergillus spp, Mucor and yeasts) were isolated from rotten yam. The organisms were paired in yam nutrient agar (YNA) and yam Sabourraud dextrose agar media. In the yam dextrose agar media (YSDA) plates, the Bacillus grew rapidly and established itself and restricted the growth of the fungi organisms, but there was no zone of inhibition. This behaviour of Bacillus on the plates of YSDA was also observed in the yams where the fungi caused rot but the rot was suppressed by the presence of the Bacillus as compared to the degree of rot observed in the control that had only spoilage fungi. The control yam showed greater rot than other yams that contained a combination of Bacillus and fungi. The t-Test analysis showed that the difference in the rot between the treated samples and the control sample is significant and this implies that the presence of Bacillus significantly reduced the growth of fungi in the samples (yams). It was revealed from this study that Bacillus subtilis treatment can be successfully used to preserve white yams in storage. Its fast growth and early establishment in the sample accounts for its antifungal strength.

Keywords: Bacillus subtilis, rot, fungi, yam

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Authors: Abeer A. Q. Ahmed, Tracey McKay

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The increasing demand for renewable fuels and chemicals is pressuring manufacturing industry toward finding more sustainable cost-effective resources. Lignocellulose, such as agro-food wastes, is a suitable equivalent to petroleum for fine chemicals and fuels production. The complex structure of lignocellulose, however, requires a variety of enzymes in order to degrade its components into their respective building blocks that can be used further for the production of various value added products. This study aimed to isolate bacterial strain with the ability to produce a variety of lignocellulosic enzymes. One bacterial isolate was identified by 16S rRNA gene sequencing and phylogenetic analysis as Bacillus safensis LCX found to have CMCase, xylanase, manganese peroxidase, lignin peroxidase, and laccase activities. The enzymes production was induced by growing Bacillus safensis LCX in solid state fermentation using wheat straw, wheat bran, and corn stover. The activities of enzymes were determined by specific colorimetric assays. This study presents Bacillus safensis LCX as a promising source for lignocellulosic enzymes. These findings can extend the knowledge on agro-food wastes valorization strategies toward a sustainable production of fuels and chemicals.

Keywords: Bacillus safensis LCX, high valued chemicals, lignocellulosic enzymes, solid state fermentation

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Authors: Vu Thu Trang, Lam Xuan Thanh, Samira Sarter, Tomoko Shimamura, Hiroaki Takeuchi　　

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

Keywords: pathogenic bacteria, aminoreductone, Maillard reaction, antimicrobial activity

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Authors: Radovan Čobanović, Milica Rankov Šicar

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The fresh cheese is in the group of perishable food which cannot be kept a long period of time. The study of sustainability have been done in order to extend the shelf-life of the product which was 15 days. According to the plan of sustainability it was defined that 35 samples had to be stored for 30 days at 2°C−6°C and analyzed every 7th day from the day of reception until 30th day. Shelf life of the cheese has expired during the study of sustainability in the period between 15th and 30th day of analyses. Cheese samples were subjected to sensory analysis (appearance, odor, taste, color, aroma) and bacteriological analyzes (Listeria monocytogenes, Salmonella spp., Bacillus cereus, Staphylococcus aureus and total plate count) according to Serbian state regulation. All analyses were tested according to ISO methodology: sensory analysis ISO 6658, Listeria monocytogenes ISO 11 290-1, Salmonella spp ISO 6579, Bacillus cereus ISO 7932, Staphylococcus aureus ISO 6888-1, and total plate count ISO 4833. Analyses showed that after fifteen days of storage at a temperature defined by the manufacturers and within the product's shelf life, the cheese did not have any noticeable changes in sensory characteristics. Smell and taste are unaffected there was no separation of whey and there was not presence of strange smell or taste. As far as microbiological analyses are concerned neither one pathogen was detected and total plate count was at level of 103 cfu/g. After expiry of shelf life in a period of 15th and 30th day of storage, the analysis showed that there was a separation of whey on the surface. Along the edge of the container was present a dried part of cheese and sour-milky smell and taste were very weakly expressed. Concerning the microbiological analyses there still were not positive results for pathogen microorganisms but the total plate count was at a level of 106cfu/g. Based on the obtained results it can be concluded that this product cannot have longer shelf life than shelf life which is already defined because there are a sensory changes that would certainly have influence on decision of customers when purchase of this product is concerned.

Keywords: sustainability, fresh cheese, shelf-life, product

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Authors: Murada Alholy, Mengshi Lin, Omar Alhaj, Mahmoud Abugoush

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Alicyclobacillus is a causative agent of spoilage in pasteurized and heat-treated apple juice products. Differentiating between this genus and the closely related Bacillus is crucially important. In this study, Fourier transform infrared spectroscopy (FT-IR) was used to identify and discriminate between four Alicyclobacillus strains and four Bacillus isolates inoculated individually into apple juice. Loading plots over the range of 1350 and 1700 cm-1 reflected the most distinctive biochemical features of Bacillus and Alicyclobacillus. Multivariate statistical methods (e.g. principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA)) were used to analyze the spectral data. Distinctive separation of spectral samples was observed. This study demonstrates that FT-IR spectroscopy in combination with multivariate analysis could serve as a rapid and effective tool for fruit juice industry to differentiate between Bacillus and Alicyclobacillus and to distinguish between species belonging to these two genera.

Keywords: alicyclobacillus, bacillus, FT-IR, spectroscopy, PCA

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Authors: Elham A. Kwildi, Yahia S. Abugnah, Nuri S. Madi

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This study was designed to evaluate the microbiological quality and safety of two types of salads prepared at a catering center affiliated with Libyan Airlines in Tripoli, Libya. Two hundred and twenty-one (221) samples (132 economy-class and 89 first- class) were used in this project which lasted for ten months. Biweekly, microbiological tests were performed which included total plate count (TPC) and total coliforms (TCF), in addition to enumeration and/or detection of some pathogenic bacteria mainly Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella sp, Listeria sp and Vibrio parahaemolyticus parahaemolyticus, By using conventional as well as compact dry methods. Results indicated that TPC of type 1 salad ranged between (<10 – 62 x 103 cfu/gm) and (<10 to 36 x103 cfu/g), while TCF were (<10 – 41 x 103 cfu/gm) and (< 10 to 66 x102 cfu/g) using both methods of detection respectively. On the other hand, TPC of type 2 salad were: (1 × 10 – 52 x 103) and (<10 – 55 x 103 cfu/gm) and in the range of (1 x10 to 45x103 cfu/g), and the (TCF) counts were between (< 10 to 55x103 cfu/g) and (< 10 to 34 x103 cfu/g) using the 1st and the 2nd methods of detection respectively. Also, the pathogens mentioned above were detected in both types of salads, but their levels varied according to the type of salad and the method of detection. The level of Staphylococcus aureus, for instance, was 17.4% using conventional method versus 14.4% using the compact dry method. Similarly, E. coli was 7.6% and 9.8%, while Salmonella sp. recorded the least percentage i.e. 3% and 3.8% with the two mentioned methods respectively. First class salads were also found to contain the same pathogens, but the level of E. coli was relatively higher in this case (14.6% and 16.9%) using conventional and compact dry methods respectively. The second rank came Staphylococcus aureus (13.5%) and (11.2%), followed by Salmonella (6.74%) and 6.70%). The least percentage was for Vibrio parahaemolyticus (4.9%) which was detected in the first class salads only. The other two pathogens Bacillus cereus and Listeria sp. were not detected in either one of the salads. Finally, it is worth mentioning that there was a significant decline in TPC and TCF counts in addition to the disappearance of pathogenic bacteria after the 6-7th month of the study which coincided with the first trial of the HACCP system at the center. The ups and downs in the counts along the early stages of the study reveal that there is a need for some important correction measures including more emphasis on training of the personnel in applying the HACCP system effectively.

Keywords: air travel, vegetable salads, foodborne outbreaks, Libya

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Authors: Abdelaziz Sheba Abdelrahman

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Compost enriched with actinomyces and Bacillus polymyxa algae as a Partial Substitute for Mineral N in Ewaise Mango Orchards Organic fertiliser, compost enriched with actinomyces, and the biofertilizer Bacillus polymyxa algae were used as a partial replacement for mineral N fertiliser in Ewaise mango orchards during the 2019 and 2020 seasons. When compared to using mineral N alone, the results showed that reducing the percentage of mineral N fertiliser from 100 to 50% and using compost enriched with actinomyces at 25 to 50% and Bacillus polymyxa had an announced promotion on leaf area, total chlorophylls, leaf N, P, and K, yield, and fruit quality. The use of compost enriched with actinomyces and Bacillus polymyxa, as well as mineral N, resulted in a significant decrease in nitrite in the pulp. Reducing mineral N to 25% of the suitable N had a negative impact on yield. The application of appropriate N via 50% inorganic N + compost enriched with actinomyces at 50% + Bacillus polymyxa algae increased yield quantitatively and qualitatively in Ewaise mango orchards. This promised treatment significantly reduced nitrite levels in the pulp fruit.

Keywords: bacillus polymyxa algae, fertiliser, biofertilizer, ewaise mango

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Authors: Priya Tomar, Pallavi Mittal

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As industrialization is inevitable and progress with rapid acceleration, the need for innovative ways to get rid of waste has increased. Recent advancement in bioresource technology paves novel ideas for recycling of factory waste that has been polluting the agro-industry, soil and water bodies. Paper industries in India are in a considerable number, where molasses and impure alcohol are still being used as raw materials for manufacturing of paper. Paper mills based on nonconventional agro residues are being encouraged due to increased demand of paper and acute shortage of forest-based raw materials. The colouring body present in the wastewater from pulp and paper mill is organic in nature and is comprised of wood extractives, tannin, resins, synthetic dyes, lignin and its degradation products formed by the action of chlorine on lignin which imparts an offensive colour to the water. These mills use different chemical process for paper manufacturing due to which lignified chemicals are released into the environment. Therefore, the chemical oxygen demand (COD) of the emanating stream is quite high. This paper presents some new techniques that were developed for the efficiency of bioremediation on paper industry. A short introduction to paper industry and a variety of presently available methods of bioremediation on paper industry and different strategies are also discussed here. For solving the above problem, two bacterial strains (Pseudomonas aeruginosa and Bacillus subtilis) and their consortia (Pseudomonas aeruginosa and Bacillus subtilis) were utilized for the pulp and paper mill effluent. Pseudomonas aeruginosa and Bacillus subtilis named as T–1, T–2, T–3, T–4, T–5, T–6, for the decolourisation of paper industry effluent. The results indicated that a maximum colour reduction is (60.5%) achieved by Pseudomonas aeruginosa and COD reduction is (88.8%) achieved by Bacillus subtilis, maximum pH changes is (4.23) achieved by Pseudomonas aeruginosa, TSS reduction is (2.09 %) achieved by Bacillus subtilis, and TDS reduction is (0.95 %) achieved by Bacillus subtilis. When the wastewater was supplemented with carbon (glucose) and nitrogen (yeast extract) source and data revealed the efficiency of Bacillus subtilis, having more with glucose than Pseudomonas aeruginosa.

Keywords: bioremediation, paper and pulp mill effluent, treated effluent, lignin

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Authors: Ali Azam Talukder, Jamsheda Ferdous Tuli, Tanzina Islam Reba, Shuvra Kanti Dey, Mamoru Yamada

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Recent global warming created by various pollutants prompted us to find new energy sources instead of fossil fuels. Fossil fuels are one of the key factors to emit various toxic gases in this planet. To solve this problem, along with the scarcity of the worldwide energy crisis, scientists are looking for various alternative options to mitigate the necessity of required future fuels. In this context, bioethanol can be one of the most suitable alternative energy sources. Bioethanol is a renewable, environment-friendly and carbon-neutral sustainable energy. In our previous study, we identified several bioethanol-producing microbes from the natural fermented sources of Bangladesh. Among them, the strain 4C encoded Saccharomyces cerevisiae produced maximum bioethanol when the fermentation temperature was 25˚C. In this study, we have established high-temperature simultaneous saccharification and fermentation process (HTSSF) by co-culturing of thermally adapted thermosensitive 4C as a fermenting agent and Bacillus amyloliquefaciens (C7), as a saccharifying agent under various physiological conditions or treatments. Conventional methods were applied for cell culture, media preparation and other experimental purposes. High-temperature adaptation of strain 4C was made from 30-42ᵒC, using either YPD or YPS media. In brief, for thermal adaptation, the temperature was periodically increased by 2ᵒC, 1ᵒC and 0.5ᵒC when medium growth temperatures were 30-36ᵒC, 36-40ᵒC, and 40-42ᵒC, respectively, where applicable. Amylase activity and bioethanol content were measured by DNS (3, 5-dinitrosalicylic acid) and solvent extraction and dichromate oxidation method, respectively. Among the various growth parameters like temperatures (30˚C, 37˚C and 42˚C), pHs (5.0, 6.0 and 7.0), carbon sources (5.0-10.0%) and ethanol stress tolerance (0.0-12.0%) etc. were tested, maximum Amylase activity (4.0 IU/ml/min) was recorded for Bacillus amyloliquefaciens (C7) at 42˚C, pH 6.0 and 10% starch. On the other hand, 4.10% bioethanol content was recorded when the thermally adapted strain 4C was co-cultured with C7 at 37ᵒC, pH 6.0 and 10.0% starch for 72 hours at HTSSF process. On the other hand, thermally non-adapted strains gave only 0.5-2.0% bioethanol content under the same physiological conditions. The thermally adapted strain 4C and strain C7, both can tolerate ethanol stress up to 12%. Altogether, a comparative study revealed that our established HTSSF process may be suitable for pilot scale and subsequently at industrial level bioethanol production.

Keywords: bioethanol, co-culture, fermentation, saccharification

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Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Patience Orobosa Olajide

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Five hydrocarbon degrading bacterial strains isolated from contaminated environment were investigated with respect to polyhydroxybutyrate (PHB) biosynthesis. Screening for bioplastic production was done on assay mineral salts agar medium containing 0.2% poly (3-hydroxybutyrate) as the sole carbon source. Two of the test bacteria were positive for PHB biosynthesis and were identified based on gram staining, biochemical tests, 16S rRNA gene sequence analysis as Pseudomonas aeruginosa and Bacillus licheniformis which grew at 37 and up to 65 °C respectively, thus suggesting the later to be thermotolerant. In this study, the effects of different carbon and nitrogen sources on PHB production in these strains were investigated. Maximum PHB production was obtained in 48 hr for the two strains and amounted to yields of 72.86 and 62.22 percentages for Bacillus licheniformis and Pseudomonas aeruginosa respectively. In these strains, glycine was the most efficient carbon sources for the production of PHB compared with other carbon (glucose, lactose, sucrose, Arabinose) and nitrogen (L- glycine, L-cysteine, DL-Tryptophan, and Potassium Nitrate) sources. The screening of microbial strains for industrial PHB production should be based on several factors including the cell’s capability to mineralize an inexpensive substrate, rate of growth and the extent of polymer accumulation.

Keywords: bacteria, poly-3-hydroxybutyrate (PHB), hydrocarbon, thermotolerant

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Authors: Han-Seul Lee, Min-Koo Kang, Sang-Ill Lee

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This study was conducted to reduce runoff by rainwater infiltration facility using attached growth with Bacillus sp., which are reported to remove nitrogen and phosphorus, as well as organic matter effectively. This study was investigated non-point pollutants removal efficiency of organic, nitrogen, and phosphorus in column using the media attached growth with Bacillus sp. To compare attached growth with bacillus sp. and detached media, two columns filled with perlite, zeolite, vermiculite, pumice, peat-moss was installed. In A column (attached growth with bacillus sp.), in case of infiltration velocity 30 mm/hr in high concentration of influent, it showed the removal efficiency (after aging term) is SS (suspended solid) 85.8±1.2 %, T-P (total phosphorus) 67.0±8.1 %, T-N (total nitrogen) 66.0±4.9 %, COD (chemical oxygen demand) 73.6±2.9 %, NH4+-N 72.7±3.0 %. In B column (detached media), in case of infiltration velocity 30 mm/hr in high concentration of influent, it showed the removal efficiency (after aging term) is SS 86.0±2.2 %, T-P 62.5±11.3 %, T-N 53.3±3.9 %, COD 34.6±3.7 %, NH4+-N 61.5±2.8 %. Removal efficiency of A column is better than B column. As the result from this study, using media with Bacillus sp. can improve an effective removal of non-point source pollutants.

Keywords: non-point source pollutants, Bacillus sp., rainwater, infiltration facility

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Authors: Min-Shiuan Liou, Yi Shan Yang, Yang-Zhan Huang, Chia-Wen Hsieh

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A high speed growing and butanol-tolerant Clostridium acetobutylicum HOL1 mutant was screened throughout continuous adaption culture with C. acetobutylicum ATCC 824. The HOL1 strain can grow well in 10 g/L butanol contained CGM medium and can produce about 12.8 g /L butanol during 24 hrs. The C. acetobutylicum HOL1 strain was able to produce 166 mM butanol with 21 mM acetone at pH 4.8, resulting in a butanol selectivity (a molar ratio of butanol to total solvents) of 0.79, which is much higher than that (0.6) of the wild-type strain C. acetobutylicum ATCC 824. The acetate and butyrate accumulation were not observed during fermentation of the HOL1 strain. A hyper-butanol producing C. acetobutylicum HOL1 (pBPHS-3), which was created to overexpress the Bacillus psychrosaccharolyticus originated specific heat-shock protein gene, hspX, from a clostridial phosphotransbutyrylase promoter, was studied for its potential to produce a high titer of butanol. Overexpression of hspX resulted in increased final butanol yield 47% and 30% higher than those of the the ATCC824 and the HOL1 strains, respectively. The remarkable high-speed growth and butanol tolerance of strain HOL1 (pBPHS-3) demonstrates that overexpression of heterogeneous stress protein-encoding gene, hspX, could help C. acetobutylicum to effectively produce a high concentration of butanol.

Keywords: Clostridium acetobutylicum, butanol, heat-shock protein, resistance

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Authors: Sahira Nsayef Muslim, Israa M. S. Al-Kadmy, Nadheema Hammood Hussein, Alaa Naseer Mohammed Ali, Buthainah Mohammed Taha, Rayim Sabah Abbood, Sarah Naji Aziz

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A novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples have the ability to produce the chitosanase enzyme when grown on chitosan substrate. Chitosanase was purified to homogeneity with a recovery yield of 35.71% and 5.5 fold of purification by using ammonium sulfate at 45% saturation followed by ion exchange chromatography on DEAE-cellulose column and gel filtration chromatography on Sephadex G-100 column. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria (biofilm producers) after using Congo Red agar and Microtiter plates methods. Highly antibiofilm of chitosanase recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation ratio to 22 and 29%, respectively compared with (100)% of control. Thus, chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug-resistant pathogen-associated infections, especially in situation where biofilms are involved.

Keywords: chitosanase, Bacillus licheniformis, vegetables, biofilm

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Authors: Jin Sung Kim, Hoon Huh

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The material properties of OFHC copper film was investigated with the High-Speed Material Micro Testing Machine (HSMMTM) at the high strain rates. The rate-dependent stress-strain curves from the experiment and the Johnson-Cook curve fitting showed large discrepancies as the plastic strain increases since the constitutive model implies no rate-dependent strain hardening effect. A new constitutive model was proposed in consideration of rate-dependent strain hardening effect. The strain rate hardening term in the new constitutive model consists of the strain rate sensitivity coefficients of the yield strength and strain hardening.

Keywords: rate dependent material properties, dynamic constitutive model, OFHC copper film, strain rate

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Authors: Rida B. Arieby, Hameed N. Hameed

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In this work, tensile tests on high density polyethylene have been carried out under various constant strain rate and strain rate jump tests. The dependency of the true stress and specially the variation of volume strain have been investigated, the volume strain due to the phenomena of damage was determined in real time during the tests by an optical extensometer called Videotraction. A modified constitutive equations, including strain rate and damage effects, are proposed, such a model is based on a non-equilibrium thermodynamic approach called (DNLR). The ability of the model to predict the complex nonlinear response of this polymer is examined by comparing the model simulation with the available experimental data, which demonstrate that this model can represent the deformation behavior of the polymer reasonably well.

Keywords: strain rate jump tests, volume strain, high density polyethylene, large strain, thermodynamics approach

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Authors: Hafiz Husnain Nawaz

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Anthracnose disease in olive, caused by the fungal pathogen Colletotrichum acutatum, is considered one of the most critical issues in olive orchards in Pakistan. This disease poses a significant threat as it results in infections that can lead to the complete damage of olive plants, affecting leaves, stems, and fruits in the field. Controlling this disease is particularly challenging due to the absence of an effective fungicide that does not pose risks to farmer health and the environment. To address this challenge, our study aimed to evaluate the antagonistic activity of a biosurfactant produced by the Bacillus subtilis PE-07 strain against the anthracnose-causing agent in olive plants. This strain was selected after screening sixty rhizobacteria strains. Additionally, we assessed the heat stability, pH range, and toxicity of the biosurfactant produced by strain PE-07. Our results revealed that the biosurfactant exhibited maximum antifungal activity against C. acutatum. In vitro studies indicated that the biosurfactant could reduce fungal activity by inhibiting the spore germination of C. acutatum. Furthermore, the biosurfactant demonstrated a wide pH and temperature range, displaying antifungal activity at pH levels ranging from 5 to 10 and a temperature range from room temperature to 110°C. To evaluate the biosurfactant's safety, we conducted toxicity tests on zebra fish (Danio rerio). The results showed that the biosurfactant had minimal harmful effects, even at maximum concentrations. In conclusion, our study confirmed that the biosurfactant produced by B. subtilis exhibited high pH and heat stability with minimal harmful effects. Therefore, it presents a promising alternative to chemical pesticides for effectively controlling olive anthracnose in Pakistan.

Keywords: biological control, heat stability and PH range, toxicity, Danio rerio

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Authors: Mantai Chen, Johnny Ching Ming Ho

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The stress-strain relationship of concrete under flexure is one of the essential parameters in assessing ultimate flexural strength capacity of RC beams. Currently, the concrete stress-strain curve in flexure is obtained by incorporating a constant scale-down factor of 0.85 in the uniaxial stress-strain curve. However, it was revealed that strain gradient would improve the maximum concrete stress under flexure and concrete stress-strain curve is strain gradient dependent. Based on the strain-gradient-dependent concrete stress-strain curve, the investigation of the combined effects of strain gradient and concrete strength on flexural strength of RC beams was extended to high strength concrete up to 100 MPa by theoretical analysis. As an extension and application of the authors’ previous study, a new flexural strength design method incorporating the combined effects of strain gradient and concrete strength is developed. A set of equivalent rectangular concrete stress block parameters is proposed and applied to produce a series of design charts showing that the flexural strength of RC beams are improved with strain gradient effect considered.

Keywords: beams, equivalent concrete stress block, flexural strength, strain gradient

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Authors: Shiv Mohan Singh, Gwyneth Matcher

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To understand the culturable microbial composition and diversity patterns, soil samples were collected from inland nunataks of Jutulsessen and Ahlmannryggen ranges in Dronning Maud Land, Antarctica. 16S rRNA, ITS and the D1/D2 domain sequencing techniques were used for characterization of microbial communities of these geographical areas. The total 37 species of bacteria such as Arthrobacter agilis, Acinetobacter baumannii, Arthrobacter flavus, Arthrobacter ginsengisoli, Arthrobacter oxydans, Arthrobacter oryzae, Arthrobacter polychromogenes, Arthrobacter sulfonivorans, Bacillus altitudinis, Bacillus cereus, Bacillus paramycoides, Brevundimonas vesicularis, Brachybacterium rhamnosum, Curtobacterium luteum, Dermacoccus nishinomiyaensis, Dietzia aerolata, Janibacter indicus, Knoellia subterranean, Kocuria palustris, Kytococcus aerolatus, Lysinibacillus sphaericus, Microbacterium phyllosphaerae, Micrococcus yunnanensis, Methylobacterium rhodesianum, Moraxella osloensis, Paracoccus acridae, Pontibacter amylolyticus, Pseudomonas hunanensis, Pseudarthrobacter siccitolerans, Pseudarthrobacter phenanthrenivorans, Rhodococcus aerolatus, Rhodococcus sovatensis, Sphingomonas daechungensis, Sphingomonas sanguinis, Stenotrophomonas pavanii, Staphylococcus gallinarum, Staphylococcus arlettae and 9 species of fungi such as Candida davisiana, Cosmospora arxii, Geomyces destructans, Lecanicillium muscarium, Memnoniella humicola, Paecilomyces lilacinus, Pseudogymnoascus verrucosus, Phaeophlebiopsis ignerii and Thyronectria caraganae were recorded. Fatty acid methyl esters (FAME) analyses of representative species of each genus have shown predominance branched and unsaturated fatty acids indicate its adaptation strategy in Antarctic cold environment. To the best of our knowledge, this is the first record of culturable bacterial communities from Jutulsessen and Ahlmannryggen ranges in Western Dronning Maud Land, Antarctica.

Keywords: antarctica, microbe, adaptation, polar

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Authors: Jiawei Chen, Jia Qu, Dianwei Ju

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316L stainless steel has good mechanical and technological properties, has been widely used in shipbuilding and aerospace manufacturing. In order to understand the effect of strain rate on the yield limit of 316L stainless steel and the constitutive relationship of the materials at different strain rates, this paper used the INSTRON-4505 electronic universal testing machine to study the mechanical properties of the tensile specimen under quasi-static conditions. Meanwhile, the Zwick-Roell RKP450 intelligent oscillometric impact tester was used to test the tensile specimens at different strain rates. Through the above two kinds of experimental researches, the relationship between the true stress-strain and the engineering stress-strain at different strain rates is obtained. The result shows that the tensile yield point of 316L stainless steel increases with the increase of strain rate, and the real stress-strain curve of the 316L stainless steel has a better normalization than that of the engineering stress-strain curve. The real stress-strain curves can be used in the practical engineering of impact stretch to improve its safety.

Keywords: impact stretch, 316L stainless steel, strain rate, real stress-strain, normalization

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Authors: Najat El-Kurdi, Sherif Hammad, Mohamed Ghazi, Sahar El-Shatoury, Khaled Zakaria

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The increasing number of plastic production and its importance to humanity in daily life made it a headache to the planet earth. The persistence of plastic wastes in the environment formed a serious problem. They are prominent with their capability to resist microbial degradation for decades. Thus, it was crucial to find ways to eliminate the plastics without depending on conventional recycling methods, which causes the formation of more hazardous compounds and doubles the problem. In this paper, mealworms were fed with a mixture of plastic wastes such as plastic bags, Styrofoam, PE foam, and plastic tarpaulins film as the sole food source for a month. Frass was collected at the end of the test and examined using FTIR analysis. Also, the gut bacteria were isolated and identified using 16S rRNA. The results show the mineralization of plastic in the frass of plastic-fed worms when compared to control. The 16S rRNA and the BLAST analysis showed that the obtained isolate belongs to the genus Bacillus Sp especially Bacillus subtilis. Phylogenetic analysis showed their relatedness to the other Bacillus species in the NCBI database.

Keywords: mealworm, waste management, plastic-degrading bacteria, gut microbiome, Bacillus sp

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Authors: Mohammad Abdel Fattah Mohammad Kewisha

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Microbial deterioration of ancient materials became one of the biggest problems facing the workers in the field of cultural heritage protection because the microbial deterioration of artifacts causes detrimental effects on the aesthetic value of the monuments due to colonization, whether they are made of inorganic materials such as stone or organic like wood, textiles, wall paintings, and paper. So, the early identification of the bacterial strains that caused deterioration is the most important point for the protection of monument objects. The present study focuses on the Bacillus spp. group, which was isolated from some biodeterioration monuments from different areas of Egypt. The investigated objects in this study were made from organic materials (cellulose), paper, textile, and wood. Isolated strains were identified up to the species level biochemically. Eleven bacterial isolates were obtained from collected samples. They were taken from different archaeological objects, four microbicides, cetrimonium bromide, sodium azide, tetraethyl ammonium bromide, and dichloroxylenol, at various concentrations ranging from 25 ppm to 500 ppm. They were screened for their antibacterial activity against the Bacillus spp. isolates, and detection of Minimum inhibitory concentration (MIC). It was also necessary to indicate the ideal Minimum inhibitory concentration for each strain for the purpose of biotreatment of the infected monuments with less damaging effect on monument materials.

Keywords: microbial deterioration, ancient materials, heritage protection, protection of monuments, biodeteriorative monuments

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Authors: Bassam O AlJohny

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The growth of Bacillus aquimaris was inhibited from 6 - 20 % of NaCl but it showed some tolerance when Diatomaceous earth (DE) added from 2 - 12% NaCl. Concerning the effect of NaCl on polyol production, we can conclude that, the test bacterium showed some tolerance to NaCl by producing glycerol up to 8 % of NaCl. Then decreased sharply. The addition of DE decrease the amount of polyol and glycerol remarkably and this due to the productive effect of DE to the bacterial cells. The SEM figures represented the presence of electron dense bodies due to the accumulation of small particles of DE as protective molecules.

Keywords: Bacillus aquimaris, Diatomaceous earth (DE), osmoticstress, sodium chloride

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Authors: Cheba Ben Amar

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Bacillus and Paenibacillus are Gram-positive aerobic endospore-forming bacteria (AEFB) and most abundant in the rhizosphere, they mediated plant growth promotion and disease protection by several complex and interrelated processes involving direct and indirect mechanisms that include nitrogen fixation, phosphate solubilization, siderophores production, phytohormones production and plant diseases control. In addition to their multiple PGPR properties, high secretory capacity, spore forming ability and spore resistance to unfavorable conditions enabling their extended commercial applications for long shelf-life. Due to these unique advantages, Bacillus species were the most an ideal candidate for developing efficient PGPR products such as biopesticides, fungicides and fertilizers. This review list all studied and reported plant growth promoting Bacillus species and strains, discuss their capacities to enhance plant growth and protection with special focusing on the most frequent species Bacillus subtilis, B. pumilus ,B. megaterium, B. amyloliquefaciens , B. licheniformis and B. sphaericus, furthermore we recapitulate the beneficial activities and mechanisms of several species and strains of the genus Paenibacillus involved in plant growth stimulation and plant disease control.

Keywords: bacillus, paenibacillus, PGPR, beneficial activities, mechanisms, growth promotion, disease control, agrobiotechnology

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Authors: Narasimhulu Korrapati

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The objective of this present study is to optimize the process parameters for batch biosorption of Cr(VI) and Cu(II) ions by Bacillus subtilis using Response Surface Methodology (RSM). Batch biosorption studies were conducted under optimum pH, temperature, biomass concentration and contact time for the removal of Cr(VI) and Cu(II) ions using Bacillus subtilis. From the studies it is noticed that the maximum biosorption of Cr(VI) and Cu(II) was by Bacillus subtilis at optimum conditions of contact time of 30 minutes, pH of 4.0, biomass concentration of 2.0 mg/mL, the temperature of 32°C in batch biosorption studies. Predicted percent biosorption of the selected heavy metal ions by the design expert software is in agreement with experimental results of percent biosorption. The percent biosorption of Cr(VI) and Cu(II) in batch studies is 80% and 78.4%, respectively.

Keywords: heavy metal ions, response surface methodology, biosorption, wastewater

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Authors: E. Soboleva, E. Sergachyova, S. G. Davydenko, T. V. Meledina

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Problem of food preservation is extremely important for mankind. Viscous damage ("illness") of bread results from development of Bacillus spp. bacteria. High temperature resistant spores of this microorganism are steady against 120°C) and remain in bread during pastries, potentially causing spoilage of the final product. Scientists are interested in further characterization of bread spoiling Bacillus spp. species. Our aim was to find weather yeast Saccharomyces cerevisiae strains that are able to produce natural antimicrobial killer factor can preserve bread illness. By diffusion method, we showed yeast antagonistic activity against spore-forming bacteria. Experimental technological parameters were the same as for bakers' yeasts production on the industrial scale. Risograph test during dough fermentation demonstrated gas production. The major finding of the study was a clear indication of the presence of killer yeast strain antagonistic activity against rope in bread causing bacteria. After demonstrating antagonistic effect of S. cerevisiae on bacteria using solid nutrient medium, we tested baked bread under provocative conditions. We also measured formation of carbon dioxide in the dough, dough-making duration and quality of the final products, when using different strains of S. cerevisiae. It is determined that the use of yeast S. cerevisiae RCAM 01730 killer strain inhibits appearance of rope in bread. Thus, natural yeast antimicrobial killer toxin, produced by some S. cerevisiae strains is an anti-rope in bread protector.

Keywords: bakers' yeasts, killer toxin, rope in bread, Saccharomyces cerevisiæ

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Authors: Mohammed H. Abu-Dieyeh, Mohammad M. Zalloum

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Plant growth-promoting rhizobacteria (PGPR) are known to enhance plant growth and/or reduce plant damage due to soil-borne pathogens. Tomato is the highest consumable vegetable world-wide including Jordan. Fusarium oxysporum is a pathogen that causes well-known damages and losses to many vegetable crops including tomato. In this study, purification of 112 isolates of PGPR strains from rhizosphere environment of different regions in Jordan was accomplished. All bacterial isolates were In-vitro screened for antagonistic effects against F. oxysporum. The eleven most effective isolates that caused 30%-50% in-vitro growth reduction of F. oxysporum were selected. 8 out of 11 of these isolates were collected from Al-Halabat (arid-land). 7 isolates of Al-Halabat exerted 40-54% In-vitro growth reduction of F. oxysporum. Four-week-old seedlings of tomato cultivar (Anjara, the most susceptible indigenous cultivar to F. oxysporum) treated with PGPR5 (Bacillus amyloliquefaciens), and exposed to F. oxysporum, showed no disease symptoms and no significant changes in biomasses or chlorophyll contents indicating a non-direct mechanism of action of PGPR on tomato plants. However PGPR3 (Bacillus sp.), PGPR4 (Bacillus cereus), and PGPR38 (Paenibacillus sp.) treated plants or PGPR treated and exposed to F. oxysporum showed a significant increasing growth of shoot and root biomasses as well as chlorophyll contents of leaves compared to control untreated plants or plants exposed to the fungus without PGPR treatment. A significant increase in number of flowers per plant was also recorded in all PGPR treated plants. The characterization of rhizobacterial strains were accomplished using 16S rRNA gene sequence analysis in addition to microscopic characterization. Further research is necessary to explore the potentiality of other collected PGPR isolates on tomato plants in addition to investigate the efficacy of the identified isolates on other plant pathogens and then finding a proper and effective methods of formulation and application of the successful isolates on selected crops.

Keywords: antagonism, arid land, growth promoting, rhizobacteria, tomato

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Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

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3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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Authors: Adebowale T. Odeyemi, Oluwafemi A. Ajenifuja

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The leachate samples collected from Erifun dumpsite along Federal Polythenic road, Ado-Ekiti, Ekiti State, were subjected to bacteriological and mineral analyses. The bacteriological estimation and isolation were done using serial dilution and pour plating techniques. Antibiotic susceptibility test was done using agar disc diffusion technique. Atomic Absorption Spectophotometry method was used to analyze the heavy metal contents in the leachate samples. The bacterial and coliform counts ranged from 4.2 × 105 CFU/ml to 2.97 × 106 CFU/ml and 5.0 × 104 CFU/ml to 2.45 x 106 CFU/ml, respectively. The isolated bacteria and percentage of occurrence include Bacillus cereus (22%), Enterobacter aerogenes (18%), Staphylococcus aureus (16%), Proteus vulgaris (14%), Escherichia coli (14%), Bacillus licheniformis (12%) and Klebsiella aerogenes (4%). The mineral value ranged as follow; iron (21.30mg/L - 25.60mg/L), zinc (1.80mg/L - 5.60mg/L), copper (1.00mg/L - 2.60mg/L), chromium (0.50mg/L - 1.30mg/L), candium (0.20mg/L - 1.30mg/L), nickel (0.20mg/L - 0.80mg/L), lead (0.05mg/L-0.30mg/L), cobalt (0.03mg/L - 0.30mg/L) and in all samples manganese was not detected. The entire organisms isolated exhibited a high level of resistance to most of the antibiotics used. There is an urgent need for awareness to be created about the present situation of the leachate in Erifun, on the need for treatment of the nearby stream and other water sources before they can be used for drinking and other domestic use. In conclusion, a good method of waste disposal is required in those communities to prevent leachate formation, percolation, and runoff into water bodies during the raining season.

Keywords: antibiotic susceptibility, dumpsite, bacteriological analysis, heavy metal

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Authors: Zhazira Shemsheyeva, Zhanara Suleimenova, Olga Shemshura, Gulnaz Mombekova, Zhanar Rakhmetova

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Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). They not only provide nutrients to the plants (direct plant growth promotion) and protect plants against the phytopathogens (indirect plant growth promotion) but also increase the soil fertility. Indirectly PGPRs improve the plant growth by becoming a biocontrol agent for a fungal pathogen. The antifungal activities of the PGPrhizobacteria were assayed against different species of phytopathogenic fungi such as Fusarium tricinctum, Fusarium oxysporum, Sclerotiniasclerotiorum, and Botrytis cinerea. Pseudomonas putidaSM-1, Azotobacter sp., and Bacillus thuringiensis AKS/16 strains have been used in experimental tests on growth inhibition of phytopathogenic fungi infecting Kidney beans. Agar well diffusion method was used in this study. Diameters of the zones of inhibition were measured in millimeters. It was found that Bacillus thuringiensis AKS/16 strain showed the lowest antifungal activity against all fungal pathogens tested. Zones of inhibition were 15-18 mm. In contrast, Pseudomonas putida SM-1 exhibited good antifungal activity against Fusarium oxysporum and Fusarium tricinctum by producing 29-30 mm clear zones of inhibition. The moderate inhibitory effect was shown by Azotobacter sp. against all fungal pathogens tested with zones of inhibition from24 to 26 mm. In summary, Pseudomonas putida SM-1 strain demonstrated the potential of controlling root rot diseases in kidney beans.

Keywords: PGPR, pseudomonas putida, kindey beans, antifungal activity

Procedia PDF Downloads 127