Search results for: bacillus cereus

Authors: Wayde Veldman, Ozlem T. Bishop, Igor Polikarpov

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In bacteria, mono and disaccharides are phosphorylated during uptake into the cell via the widely used phosphoenolpyruvate (PEP)-dependent phosphotransferase transport system. As an initial step in the phosphorylated disaccharide metabolism pathway, certain glycoside hydrolase family 1 (GH1) enzymes play a crucial role in releasing phosphorylated and non-phosphorylated monosaccharides. However, structural determinants for the specificity of these enzymes still need to be clarified. GH1 enzymes are known to have a wide array of functions. According to the CAZy database, there are twenty-one different enzymatic activities in the GH1 family. Here, the structure and substrate specificity of a GH1 enzyme from Bacillus licheniformis, hereafter known as BlBglH, was investigated. The sequence of the enzyme BlBglH was compared to the sequences of other characterized GH1 enzymes using sequence alignment, sequence identity calculations, phylogenetic analysis, and motif discovery. Through these various analyses, BlBglH was found to have sequence features characteristic of the 6-phospho-β-glucosidase activity enzymes. Additionally, motif and structure comparisons of the three most commonly studied GH1 enzyme-activities revealed a shared loop amongst the different structures that consist of different sequence motifs – this loop is thought to guide specific substrates (depending on activity) towards the active-site. To further affirm BlBglH enzyme activity, molecular docking and molecular dynamics simulations were performed. Docking was carried out using 6-phospho-β-glucosidase enzyme-activity positive (p-Nitrophenyl-beta-D-glucoside-6-phosphate) and negative (p-Nitrophenyl-beta-D-galactoside-6-phosphate) control ligands, followed by 400 ns molecular dynamics simulations. The positive-control ligand maintained favourable interactions within the active site until the end of the simulation. The negative-control ligand was observed exiting the enzyme at 287 ns. Binding free energy calculations showed that the positive-control complex had a substantially more favourable binding energy compared to the negative-control complex. Jointly, the findings of this study suggest that the BlBglH enzyme possesses 6-phospho-β-glucosidase enzymatic activity.

Keywords: 6-P-β-glucosidase, glycoside hydrolase 1, molecular dynamics, sequence analysis, substrate specificity

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Authors: Evgenia Iakovleva, Juha Koivisto, Pasi Karppinen, J. Inkinen, Mikko Alava

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Preventing the spread of infectious diseases throughout the worldwide is one of the most important tasks of modern health care. Infectious diseases not only account for one fifth of the deaths in the world, but also cause many pathological complications for the human health. Touch surfaces pose an important vector for the spread of infections by varying microorganisms, including antimicrobial resistant organisms. Further, antimicrobial resistance is reply of bacteria to the overused or inappropriate used of antibiotics everywhere. The biggest challenges in bacterial detection by existing methods are non-direct determination, long time of analysis, the sample preparation, use of chemicals and expensive equipment, and availability of qualified specialists. Therefore, a high-performance, rapid, real-time detection is demanded in rapid practical bacterial detection and to control the epidemiological hazard. Among the known methods for determining bacteria on the surfaces, Hyperspectral methods can be used as direct and rapid methods for microorganism detection on different kind of surfaces based on fluorescence without sampling, sample preparation and chemicals. The aim of this study was to assess the relevance of such systems to remote sensing of surfaces for microorganisms detection to prevent a global spread of infectious diseases. Bacillus subtilis and Escherichia coli with different concentrations (from 0 to 10x8 cell/100µL) were detected with hyperspectral camera using different filters as visible visualization of bacteria and background spots on the steel plate. A method of internal standards was applied for monitoring the correctness of the analysis results. Distances from sample to hyperspectral camera and light source are 25 cm and 40 cm, respectively. Each sample is optically imaged from the surface by hyperspectral imaging system, utilizing a JAI CM-140GE-UV camera. Light source is BeamZ FLATPAR DMX Tri-light, 3W tri-colour LEDs (red, blue and green). Light colors are changed through DMX USB Pro interface. The developed system was calibrated following a standard procedure of setting exposure and focused for light with λ=525 nm. The filter is ThorLabs KuriousTM hyperspectral filter controller with wavelengths from 420 to 720 nm. All data collection, pro-processing and multivariate analysis was performed using LabVIEW and Python software. The studied human eye visible and invisible bacterial stains clustered apart from a reference steel material by clustering analysis using different light sources and filter wavelengths. The calculation of random and systematic errors of the analysis results proved the applicability of the method in real conditions. Validation experiments have been carried out with photometry and ATP swab-test. The lower detection limit of developed method is several orders of magnitude lower than for both validation methods. All parameters of the experiments were the same, except for the light. Hyperspectral imaging method allows to separate not only bacteria and surfaces, but also different types of bacteria, such as Gram-negative Escherichia coli and Gram-positive Bacillus subtilis. Developed method allows skipping the sample preparation and the use of chemicals, unlike all other microbiological methods. The time of analysis with novel hyperspectral system is a few seconds, which is innovative in the field of microbiological tests.

Keywords: Escherichia coli, Bacillus subtilis, hyperspectral imaging, microorganisms detection

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Authors: Vanja Vlajkov, Ivana PajčIn, Mila Grahovac, Marta Loc, Dragana Budakov, Jovana Grahovac

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The rhizosphere soil refers to the plant roots' dynamic environment characterized by their inhabitants' high biological activity. Rhizospheric bacteria are recognized as effective biocontrol agents and considered cardinal in alternative strategies for securing ecological plant diseases management. The need to suppress fungal pathogens is an urgent task, not only because of the direct economic losses caused by infection but also due to their ability to produce mycotoxins with harmful effects on human health. Aspergillus and Fusarium species are well-known producers of toxigenic metabolites with a high capacity to colonize crops and enter the food chain. The bacteria belonging to the Bacillus genus has been conceded as a plant beneficial species in agricultural practice and identified as plant growth-promoting rhizobacteria (PGPR). Besides incontestable potential, the full commercialization of microbial biopesticides is in the preliminary phase. Thus, there is a constant need for estimating the suitability of novel strains to be used as a central point of viable bioprocess leading to market-ready product development. In the present study, 76 potential producing strains were isolated from the rhizosphere soil, sampled from different localities in the Autonomous Province of Vojvodina, Republic of Serbia. The selective isolation process of strains started by resuspending 1 g of soil samples in 9 ml of saline and incubating at 28° C for 15 minutes at 150 rpm. After homogenization, thermal treatment at 100° C for 7 minutes was performed. Dilution series (10-1-10-3) were prepared, and 500 µl of each was inoculated on nutrient agar plates and incubated at 28° C for 48 h. The pure cultures of morphologically different strains indicating belonging to the Bacillus genus were obtained by the spread-plate technique. The cultivation of the isolated strains was carried out in an Erlenmeyer flask for 96 h, at 28 °C, 170 rpm. The antagonistic activity screening included two phytopathogenic fungi as test microorganisms: Aspergillus sp. and Fusarium sp. The mycelial growth inhibition was estimated based on the antimicrobial activity testing of cultivation broth by the diffusion method. For the Aspergillus sp., the highest antifungal activity was recorded for the isolates Kro-4a and Mah-1a. In contrast, for the Fusarium sp., following 15 isolates exhibited the highest antagonistic effect Par-1, Par-2, Par-3, Par-4, Kup-4, Paš-1b, Pap-3, Kro-2, Kro-3a, Kro-3b, Kra-1a, Kra-1b, Šar-1, Šar-2b and Šar-4. One-way ANOVA was performed to determine the antagonists' effect statistical significance on inhibition zone diameter. Duncan's multiple range test was conducted to define homogenous groups of antagonists with the same level of statistical significance regarding their effect on antimicrobial activity of the tested cultivation broth against tested pathogens. The study results have pointed out the significant in vitro potential of the isolated strains to be used as biocontrol agents for the suppression of the tested mycotoxigenic fungi. Further research should include the identification and detailed characterization of the most promising isolates and mode of action of the selected strains as biocontrol agents. The following research should also involve bioprocess optimization steps to fully reach the selected strains' potential as microbial biopesticides and design cost-effective biotechnological production.

Keywords: Bacillus, biocontrol, bioprocess, mycotoxigenic fungi

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Authors: Muhammad Arshad, M. Sufian, Muhammad D. Gogi, A. Aslam

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The transgenic cotton expressing Bacillus thuringiensis (Bt) provides an effective control of Helicoverpa armigera, a most damaging pest of the cotton crop. However, Bt cotton may not be the optimal solution owing to the selection pressure of Cry toxins. As Bt cotton express the insecticidal proteins throughout the growing seasons, there are the chances of resistance development in the target pests. A regular monitoring and surveillance of target pest’s baseline susceptibility to Bt Cry toxins is crucial for early detection of any resistance development. The present study was conducted to monitor the changes in the baseline susceptibility of the field population of H. armigera to Bt Cry1Ac toxin. The field-collected larval populations were maintained in the laboratory on artificial diet and F1 generation larvae were used for diet incorporated diagnostic studies. The LC₅₀ and MIC₅₀ were calculated to measure the level of resistance of population as a ratio over susceptible population. The monitoring results indicated a significant difference in the susceptibility (LC₅₀) of H. armigera for first, second, third and fourth instar larval populations sampled from different cotton growing areas over the study period 2016-17. The variations in susceptibility among the tested insects depended on the age of the insect and susceptibility decreased with the age of larvae. The overall results show that the average resistant ratio (RR) of all field-collected populations (FSD, SWL, MLT, BWP and DGK) exposed to Bt toxin Cry1Ac ranged from 3.381-fold to 7.381-fold for 1st instar, 2.370-fold to 3.739-fold for 2nd instar, 1.115-fold to 1.762-fold for 3rd instar and 1.141-fold to 2.504-fold for 4th instar, depicting maximum RR from MLT population, whereas minimum RR for FSD and SWL population. The results regarding moult inhibitory concentration of H. armigera larvae (1-4th instars) exposed to different concentrations of Bt Cry1Ac toxin indicated that among all field populations, overall Multan (MLT) and Bahawalpur (BWP) populations showed higher MIC₅₀ values as compared to Faisalabad (FSD) and Sahiwal (SWL), whereas DG Khan (DGK) population showed an intermediate moult inhibitory concentrations. This information is important for the development of more effective resistance monitoring programs. The development of Bt Cry toxins baseline susceptibility data before the widespread commercial release of transgenic Bt cotton cultivars in Pakistan is important for the development of more effective resistance monitoring programs to identify the resistant H. armigera populations.

Keywords: Bt cotton, baseline, Cry1Ac toxins, H. armigera

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Authors: Nassima Behidj-Benyounes, Nadjiba Chebouti, Thoraya Dahmane, Amina Henni

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This work examines the study of the antimicrobrial effect of oil extracted from the seeds of Argania spinosa L. (Sapotaceae) in the area of Stida (Mostaganem). This natural substance is extracted by using the Soxhlet. The antimicrobial activity of this oil is evaluated on several microorganisms. It has been tested on five bacterial strains; Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus subtilis and Staphylococcus aureus. The extract has been studied by using Candida albicans. It should be noted that these agents are characterized by a high frequency of contamination and pathogenicity. Through this study, we note that these microorganisms are moderately sensitive to the argan oil.

Keywords: Argania spinosa, oil, several microorganisms, almonds, antimicrobial activity

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Authors: Asim Abbasi, Muhammad Sufyan, Iqra, Hafiza Javaria Ashraf

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The share of vector-borne diseases (VBDs) in the global burden of infectious diseases is almost 17%. The advent of new drugs and latest research in medical science helped mankind to compete with these lethal diseases but still diseases transmitted by different mosquito species, including filariasis, malaria, viral encephalitis and dengue are serious threats for people living in disease endemic areas. Injudicious and repeated use of pesticides posed selection pressure on mosquitoes leading to development of resistance. Hence biological control agents are under serious consideration of scientific community to be used in vector control programmes. Fish have a history of predating immature stages of different aquatic insects including mosquitoes. The noteworthy examples in Africa and Asia includes, Aphanius discolour and a fish in the Panchax group. Moreover, common mosquito fish, Gambusia affinis predates mostly on temporary water mosquitoes like anopheline as compared to permanent water breeders like culicines. Mosquitoes belonging to genus Toxorhynchites have a worldwide distribution and are mostly associated with the predation of other mosquito larvae habituating with them in natural and artificial water containers. These species are harmless to humans as their adults do not suck human blood but feeds on floral nectar. However, their activity is mostly temperature dependent as Toxorhynchites brevipalpis consume 359 Aedes aegypti larvae at 30-32 ºC in contrast to 154 larvae at 20-26 ºC. Although many bacterial species were isolated from mosquito cadavers but those belonging to genus Bacillus are found highly pathogenic against them. The successful species of this genus include Bacillus thuringiensis and Bacillus sphaericus. The prime targets of B. thuringiensis are mostly the immatures of genus Aedes, Culex, Anopheles and Psorophora while B. sphaericus is specifically toxic against species of Culex, Psorophora and Culiseta. The entomopathogenic nematodes belonging to family, mermithidae are also pathogenic to different mosquito species. Eighty different species of mosquitoes including Anopheles, Aedes and Culex proved to be highly vulnerable to the attack of two mermithid species, Romanomermis culicivorax and R. iyengari. Cytoplasmic polyhedrosis virus was the first described pathogenic virus, isolated from the cadavers of mosquito specie, Culex tarsalis. Other viruses which are pathogenic to culicine includes, iridoviruses, cytopolyhedrosis viruses, entomopoxviruses and parvoviruses. Protozoa species belonging to division microsporidia are the common pathogenic protozoans in mosquito populations which kill their host by the chronic effects of parasitism. Moreover, due to their wide prevalence in anopheline mosquitoes and transversal and horizontal transmission from infected to healthy host, microsporidia of the genera Nosema and Amblyospora have received much attention in various mosquito control programmes. Fungal based mycopesticides are used in biological control of insect pests with 47 species reported virulent against different stages of mosquitoes. These include both aquatic fungi i.e. species of Coelomomyces, Lagenidium giganteum and Culicinomyces clavosporus, and the terrestrial fungi Metarhizium anisopliae and Beauveria bassiana. Hence, it was concluded that the integrated use of all these biological control agents can be a healthy contribution in mosquito control programmes and become a dire need of the time to avoid repeated use of pesticides.

Keywords: entomopathogenic nematodes, protozoa, Toxorhynchites, vector-borne

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Authors: Messaouda Belaid, Salima Kebbouche-Gana

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The negative effect of the Varroa bee colony is very important. They cause morphological and physiological changes, causing a decrease in performance of individuals and long-term death of the colony. Indirectly, they weaken the bees become much more sensitive to the different pathogenic organisms naturally present in the colony. This work aims to research secondary infections of microbial origin occurred in the worker bee nurse due to parasitism by Varroa destructor. The feeding behaviour of Varroa may causes damaging host integument. The results show that the microbial contamination enable to be transmitted into honeybee heamocoel are Bacillus sp, Pseudomonas sp, Enterobacter, Aspergillus.

Keywords: honeybee, Apis mellifera intermissa, microbial contamination, Varroa destructor

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Authors: F. Madani, M. Doudi, L. Rahimzadeh Torabi

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The irregular consumption of current antibiotics contributes to an escalation in antibiotic resistance among urinary pathogens on a global scale. The objective of this research was to investigate the process of biologically synthesized silver nanoparticles through the utilization of Zataria multiflora extract. Additionally, the study aimed to evaluate the efficacy of these synthesized nanoparticles in inhibiting the growth of multi-drug resistant negative bacillus bacteria, which commonly contribute to urinary tract infections. The botanical specimen utilized in the current research investigation was Z. multiflora, and its extract was produced employing the Soxhlet extraction technique. The study examined the green synthesis conditions of silver nanoparticles by considering three key parameters: the quantity of extract used, the concentration of silver nitrate salt, and the temperature. The particle dimensions were ascertained using the Zetasizer technique. In order to identify synthesized Silver nanoparticles TEM, XRD, and FTIR methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through a biological method, different concentrations of silver nanoparticles were studied on 140 cases of Multiple drug resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections, for identification of bacteria were used of PCR test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were subjected to analysis using the statistical software SPSS, specifically employing nonparametric Kruskal-Wallis and Mann-Whitney tests. This study yielded noteworthy findings regarding the impacts of varying concentrations of silver nitrate, different quantities of Z. multiflora extract, and levels of temperature on nanoparticles. Specifically, it was observed that an increase in the concentration of silver nitrate, extract amount, and temperature resulted in a reduction in the size of the nanoparticles synthesized. However, the impact of the aforementioned factors on the index of particle diffusion was found to be statistically non-significant. According to the transmission electron microscopy (TEM) findings, the particles exhibited predominantly spherical morphology, with a diameter spanning from 25 to 50 nanometers. Nanoparticles in the examined sample. Nanocrystals of silver. FTIR method illustrated that the spectrums of Z. multiflora and synthesized nanoparticles had clear peaks in the ranges of 1500-2000, and 3500 - 4000. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E. coli, A. bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125 mg/ml and for A. bumanii 250 mg/ml. Comparing the growth inhibitory effect of chemically synthesized the results obtained from the experiment indicated that both nanoparticles and biologically synthesized nanoparticles exhibit a notable growth inhibition effect. Specifically, the chemical method of synthesizing nanoparticles demonstrated the highest level of growth inhibition at a concentration of 62.5 mg/mL The present study demonstrated an inhibitory effect on bacterial growth, facilitating the causative factors of urine infection and multidrug resistance (MDR).

Keywords: multiple drug resistance, negative bacillus bacteria, urine infection, Zataria multiflora

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Authors: Fatma Acheuk, Akila Hamichi, Siham Semmar

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The crude ethanolic extract from Solenostemma argel was obtained by maceration of leaves and stems of the plant. Phytochimical study revealed the richness of the species on flavonoids, alkaloids, tannins and glycosides. Antimicrobial activity of the growth of clinical isolates of Eschirichia coli, Pseudomonas aeriginosa, Staphylococus aureus and Bacillus Subtilis was carried out using agar disc diffusion. The results of the study revealed that the test compound has antimicrobial activity against gram-positive bacteria which are resistant to commonly antimicrobial agents used. However, no effect was observed on other species tested.

Keywords: Solenostemma argel, crude extract, phytochemical screening, antimicrobial activity

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Authors: Ibrahim Isah Lakan, Nasiru Ibrahim

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These Phytochemical screening, Antimicrobial activities and mineral Determination of aqueous extract of Stachtarpheta indica were assessed. The result reveals the presence of flavonoids, tannins, saponins, alkaloids, glycosides and anthraquinones. The disc diffusion of aqueous extract showed Escherichia coli, 13 and antibiotic, 19 mm; Bacillus subtilis, 10 and anti –biotic, 17 mm; Klebsiller pnemuoniae , 14 and antibiotic, 24mm and Pseudmonas aeruginosa, 24 and antibiotic, 36 mm which are all comparable with the standard antibiotic cyprotomycin. The mineral content determination by flame photometer revealed that 1.25 (Na+), 0.85 (K +), 1.75 (Ca 2+) % which is a clear indication of the safety of the extract for the hypertensive patients and could be used to lower blood pressure.

Keywords: microbials, mineral, phytochemicals, stachtarpheta indica extracts

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Authors: Assel E. Molzhigitova, Amankeldi K. Sadanov, Elvira T. Ismailova, Kulyash A. Iskandarova, Olga N. Shemshura, Ainur I. Seitbattalova

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Fire blight is a very harmful for commercial apple and pear production quarantine bacterial disease. To date, several different methods have been proposed for disease control, including the use of copperbased preparations and antibiotics, which are not always reliable or effective. The use of bacteria as biocontrol agents is one of the most promising and eco-friendly alternative methods. Bacteria with protective activity against the causal agent of fire blight are often present among the epiphytic microorganisms of the phyllosphere of host plants. Therefore, the main objective of our study was screening of local epiphytic bacteria as possible antagonists against Erwinia amylovora, the causal agent of fire blight. Samples of infected organs of apple and pear trees (shoots, leaves, fruits) were collected from the industrial horticulture areas in various agro-ecological zones of Kazakhstan. Epiphytic microorganisms were isolated by standard and modified methods on specific nutrient media. The primary screening of selected microorganisms under laboratory conditions to determine the ability to suppress the growth of Erwinia amylovora was performed by agar-diffusion-test. Among 142 bacteria isolated from the fire blight host plants, 5 isolates, belonging to the genera Bacillus, Lactobacillus, Pseudomonas, Paenibacillus and Pantoea showed higher antagonistic activity against the pathogen. The diameters of inhibition zone have been depended on the species and ranged from 10 mm to 48 mm. The maximum diameter of inhibition zone (48 mm) was exhibited by B. amyloliquefaciens. Less inhibitory effect was showed by Pantoea agglomerans PA1 (19 mm). The study of inhibitory effect of Lactobacillus species against E. amylovora showed that among 7 isolates tested only one (Lactobacillus plantarum 17M) demonstrated inhibitory zone (30 mm). In summary, this study was devoted to detect the beneficial epiphytic bacteria from plants organs of pear and apple trees due to fire blight control in Kazakhstan. Results obtained from the in vitro experiments showed that the most efficient bacterial isolates are Lactobacillus plantarum 17M, Bacillus amyloliquefaciens MB40, and Pantoea agglomerans PA1. These antagonists are suitable for development as biocontrol agents for fire blight control. Their efficacies will be evaluated additionally, in biological tests under in vitro and field conditions during our further study.

Keywords: antagonists, epiphytic bacteria, Erwinia amylovora, fire blight

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Authors: Aya Tanaka, Mariko Era, Shiho Sakai, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Aspergillus niger and Aspergillus oryzae are used as koji fungi in the spot of the brewing. Since koji-muro (room for making koji) was a low level of airtightness, microbial contamination has long been a concern to the alcoholic beverage production. Therefore, we focused on the fatty acid salt which is the main component of soap. Fatty acid salts have been reported to show some antibacterial and antifungal activity. So this study examined antimicrobial activities against Aspergillus and Bacillus spp. This study aimed to find the effectiveness of the fatty acid salt in koji-muro as antimicrobial agents. Materials & Methods: A. niger NBRC 31628, A. oryzae NBRC 5238, A. oryzae (Akita Konno store) and Bacillus subtilis NBRC 3335 were chosen as tested. Nine fatty acid salts including potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K) and linolenate (C18:3K) at 350 mM and pH 10.5 were used as antimicrobial activity. FASs and spore suspension were prepared in plastic tubes. The spore suspension of each fungus (3.0×104 spores/mL) or the bacterial suspension (3.0×105 CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). The mixtures were incubated at 25 ℃. Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 ℃. The MIC (minimum inhibitory concentration) is defined as the lowest concentration of drug sufficient for inhibiting visible growth of spore after 10 min of incubation. MICs against fungi and bacteria were determined using the two-fold dilution method. Each fatty acid salt was separately inoculated with 400 µL of Aspergillus spp. or B. subtilis NBRC 3335 at 3.0 × 104 spores/mL or 3.0 × 105 CFU/mL. Results: No obvious change was observed in tested fatty acid salts against A. niger and A. oryzae. However, C12K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. Thus, C12K suppressed 99.999 % of bacterial growth. Besides, C10K was the antibacterial effect of 5 log-unit incubated time for 180 min against B. subtilis. C18:1K, C18:2K and C18:3K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. However, compared to saturated fatty acid salts to unsaturated fatty acid salts, saturated fatty acid salts are lower cost. These results suggest C12K has potential in the field of koji-muro. It is necessary to evaluate the antimicrobial activity against other fungi and bacteria, in the future.

Keywords: Aspergillus, antimicrobial, fatty acid salts, koji-muro

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Authors: M. N. Alo, U. C. C. Egbule, J. O. Orji, C. J. Aneke

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Microbiological analysis of soil from Onu-Ebonyi Izzi local government area of Ebonyi State, Nigeria contaminated with inorganic fertilizer was carried out with a view to determine the effect of the fertilizer on the microbial flora of the soil. soil samples were analyzed for microbial burden. the result showed that the following organisms were isolated with their frequency of their occurrence as follows:pseudomonas species (33.3%) and aspergillus species (54.4%) had the highest frequncy of occurence in the whole sample of batches, while streptococcus species had 6.0% and Geotrichum species (5.3%) had the least and other predominant microorganism isolated: bacillus species,staphylococcus species and vibrio species, Escherichia species, rhzizopus species, mucor species and fusaruim species. From the result, it could be concluded that the soil was contaminated and this could affect adversely the fertility of the soil .

Keywords: soil, bacteria, fungi, inorganic fertilizer, Onu- Ebonyi

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Authors: Fatimah M. Alshehrei

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For many years, synthetic pesticides have been used to kill pests; due to their toxicity and pollution, they are now a risk to human and environmental health. Lately, biopesticides have emerged as possible substitutes for petrochemical pesticides. The sources of biopesticides are widely accessible, easily biodegradable, have a variety of modes of action, are less expensive, and have little toxicity toward humans and other creatures that aren't the intended targets. Plants, bacteria, and insects are used to create biopesticides, they used in controlling diseases in crops. Microbial pesticides are produced from different microorganisms such as Trichoderma, Bacillus, Pseudomonas, and Beauveria. Also, botanical pesticides have already been commercialized; they are extracted from neem, pyrethrum, azadirachtin, etc. This paper describes biopesticide categories, their sources, mode of action, advantages and disadvantages, and their role in sustainable agriculture.

Keywords: biopesticides categories, formulation, mode of action, pest control

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Authors: Abdol-Hassan Doulah

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In this study, antimicrobial activity of essential oil and ethyl acetate and ether extracts of S. nemorosa were examined against some species of bacteria and fungi. The essential oil of the aerial part of S. nemorosa was examined by GC and GC-MS. In the essential oil of S. nemorosa 26 Compounds have been identified. 2-Nonanone (44.09 %), 2-Undecanone (33.79 %), E-Caryophyllene (3.74 %) and 2-Decanone (2.89 %) were the main components of the essential oil. The essential oil analysis showed greatest antimicrobial activity against Staphylococcus epidermidis (5.3 μg/ml) and S. cerevisiae (9.3 μg/ml). The ethyl acetate showed greatest antimicrobial activity against Bacillus subtilis (106.7 μg/ml), Candida albicans (5.3 μg/ml) and ether extract showed greatest antimicrobial activity against Klebseilla pneumoniae (10.7 μg/ml) and Saccharomyces cerevisiae (10.7 μg/ml). In conclusion, we suggest that the antimicrobial activity of S. nemorosa may be due to its content of germacrene and linalool.

Keywords: antibacterial activity, antifungal activity, Salvia nemorosa L., essential oils, biological activity

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Authors: G. A. Amin, A. D. Al-Talhi

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The bacterium B. subtilis produced surfactin in conventional batch culture as a growth associated product and a growth rate (0.4 h-1). A fed-batch process was developed and the fermentative substrate and other nutrients were fed on hourly basis and according to the growth rate of the bacterium. Conversion of different quantities of Maldex-15 into surfactin was investigated in five different fermentation runs. In all runs, most of Maldex-15 was consumed and converted into surfactin and cell biomass with appreciable efficiencies. The best results were obtained with fermentation run supplied with 200 g Maldex-15. Up to 35.4 g.l-1 of surfactin and cell biomass of 30.2 g.l-1 were achieved in 12 hrs. Also, markedly substrate yield of 0.269 g/g and volumetric reactor productivity of 2.61 g.1-1.h-1 were obtained confirming the establishment of a cost effective commercial surfactin production.

Keywords: Bacillus subtilis, biosurfactant, exponentially fed-batch fermentation, surfactin

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Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej

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Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.

Keywords: sugarcane bagasse, microorganisms, pretreatment, enzymatic hydrolysis

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Authors: Ajuna B. Henry

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In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

Procedia PDF Downloads 19

Authors: Uftah Ali M. Shushni, Viola Stuppec, Ulrike Lindequist

Abstract:

Because of the evolving resistance of microorganisms to existing antibiotics, there is an increasing need for new antibiotics not only in human but also in veterinary medicine. As part of our ongoing work on the secondary metabolites produced by marine fungi, the organic extract of the culture filtrate of an Ascomycetous fungus, which was found on driftwood collected from the coast of the Greifswalder Bodden, Baltic Sea, Germany displayed antimicrobial activity against some fish and human pathogenic bacteria. Bioactivity-guided column chromatographic separation led to the isolation of 6-Deoxybostrycoidin. The structure was determined from the interpretation of spectroscopic data (UV, MS, and NMR). 6-Deoxybostrycoidin exhibited in vitro activity against Bacillus subtilis, Staphylococcus aureus and Flexibacter maritimus with minimal inhibitory concentrations of 25, 12.5 and 12.5 μg/ml respectively.

Keywords: marine fungi, fish pathogenic bacteria, microorganism, medicine

Procedia PDF Downloads 529

Authors: S. M. Abd-Altwab, M. R. Noor El-Din

Abstract:

This paper aims to the synthesis of new ethoxylated amide as bactericides to prevent the growth of Gram +ve and –ve bacteria of water-in-diesel fuel nanoemulsions over a long period of time as three months. To realize it, eight kinetically stable water-in-diesel fuel nanoemulsions differing in surfactant concentrations and water contents ranging from 4 to 8 and 5 to 8 wt.,wt.,% of total weight of the nanoemulsions, respectively were formed at a temperature of 20 °C. The performance of this ethoxylated amide as bactericides agents against two strains of Gram-negative bacteria, namely, Pseudomonas aeruginosa and Escherichia coli, and two strains of Gram-positive bacteria namely, Staphylococcus aureus and Bacillus subtilis, were evaluated as antimicrobial agents. The maximum and minimum antimicrobial activities were 85 and 71 % against S. aureus and E. coli, respectively, at a concentration of 5 mg/l, pH 7, and 37 °C.

Keywords: nanoemulsion, bacteriocide, diesel fuel, emulsifier

Procedia PDF Downloads 363

Authors: Manfreda Gerardo, Pasquali Frederique, Pepe Tiziana, Anastasio Aniello, Ianieri Adriana

Abstract:

The aim of the research was to evaluate the shelf life of a REPFED’s product (lasagna alla bolognese), developed as a product to be marketed fresh after defrosting. Three different samples were prepared: A, B and C, which presented differences in relation to the recipe, pasteurization technique and packaging on which the trend of the shelf-life indicator parameters was evaluated during a period of prolonged shelf life. The analytical plan involved the measurement of microbiological, chemical-physical and organoleptic parameters over 7 moments of storage selected in a period of 33 days. CBT, LAB, enterobacteria, E. coli, yeasts, molds, S. coagulase positive, B. cereus, Salmonella spp and L. monocytogenes, pH, Aw, Kreiss test, peroxides, atmosphere inside the packages, and organoleptic characteristics were determined. The results demonstrated the effect of post-packaging pasteurization on the shelf life of fresh from frozen products. However, the products pasteurized at 95°C in the absence of steam showed microbiological parameters that were not appropriate for an extended shelf life of up to 60 days. On the contrary, the samples pasteurized at 98°C with steam saturation and counterpressure showed values compatible with an extended shelf life. The results of the chemical-physical analyses highlighted how recipe and packaging affect the chemical-physical and organoleptic parameters. In conclusion, this preliminary study confirmed the effectiveness of post-packaging pasteurization treatments aimed at extending the shelf life of the product, helping the food company to occupy market niches even very distant from the production sites.

Keywords: shelf life, REPFED’s product, extended durability, pasteurization

Procedia PDF Downloads 28

Authors: Suchao Donpudsa, Anchalee Tassanakajon, Vichien Rimphanitchayakit

Abstract:

A crustin gene, LV-SWD1, previously found in the hemocyte cDNA library of Litopenaeus vannamei, contains the open reading frames of 288 bp encoding a putative protein of 96 amino acid residues. The putative signal peptides of the LV-SWD1 were identified using the online SignalP 3.0 with predicted cleavage sites between Ala24-Val25, resulting in 72 residue mature protein with calculated molecular mass of 7.4 kDa and predicted pI of 8.5. This crustin contains a Arg-Pro rich region at the amino-terminus and a single whey acidic protein (WAP) domain at the carboxyl-terminus. In order to characterize their properties and biological activities, the recombinant crustin protein was produced in the Escherichia coli expression system. Antimicrobial assays showed that the growth of Bacillus subtilis was inhibited by this recombinant crustin with MIC of about 25-50 µM.

Keywords: crustin, single whey acidic protein, Litopenaeus vannamei, antimicrobial activity

Procedia PDF Downloads 244

Authors: I. L. Ibrahim, A. Mann, B. M. Abdullahi

Abstract:

Infectious diseases are prevalent in developing countries and plant extracts are known to contained bioactive compounds that can be used in the management of these diseases. The entire plant of Chrysanthellum indicum (Linn) was air-dried and pulverized into fine powder and then percolated to give ethanol and aqueous extracts. These extracts were phytochemically screened for metabolites and evaluated antibacterial activity against some pathogenic organisms Klebsilla, pneumonia, Bacillus subtilis, and Pseudomonas aeruginosa using agar dilution method. It was found that crude extracts of C. indicum revealed the presence of saponins, tannins, alkaloids, steroidal nucleus, cardiac glycosides, and coumarin while flavonoids and anthraquinones were absent. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the active extract of C. indicum shows that the extract could be a potential source of antibacterial agents.

Keywords: antibacterial activity, Chrysanthellum indicum, infectious diseases, phytochemical screening

Procedia PDF Downloads 525

Authors: Aqeel Ahmed Shad, Tanveer Ahmad, Muhammad Farooq Iqbal, Muhammad Javaid Asad

Abstract:

The supplemental effects of novel protease produced from Bacillus subtilis K-5 and beta-mannanase were evaluated on growth performance, carcass characteristics, nutrients digestibility, blood profile and intestinal morphometry of broilers fed guar meal (Cyamopsis tetragonoloba) based diets with reduced Crude Protein (CP), Essential Amino Acids (EAAs), and Metabolizable energy (ME) contents. One-day old Ross 308 broiler chicks (n=360) were randomly allotted to thirty six experimental units in a way that each of the nine dietary treatments received four replicates with ten birds per replicate. A control diet without guar meal (0GM) was formulated with standard nutrient specifications of Ross 308 for the starter and finisher phases. Two negative control diets, one with 5% (5GM) and second with 10% (10GM) guar meal, were formulated with reduction of 5% CP, 5% EAAs and 80 Kcal/kg ME. These three basal diets (no enzyme) were supplemented with novel protease enzyme (PROT) and commercial beta-mannanase (Beta-M) enzyme. The birds were reared up to 35d of age. The data on weekly body weight gain (BWG) and feed intake were recorded to compute feed:gain for the starter (0-21d) and finisher (22-35d) phases. At the end of 35d of experimental period, four birds per experimental unit were randomly selected for blood samples collection and later slaughtered for ileal digesta, intestinal tract and carcass trait sampling. The data on overall performance (1-35d) indicated improved (P<0.05) BWG and feed:gain in birds supplemented with PROT (1.41% and 1.67) and Beta-M (2.79% and 1.64) than non-supplemented groups. Improved (P<0.05) carcass yield, breast meat yield and thigh meat yield were noted with the supplementation of Beta-M. However, non-significant (P>0.05) effect on carcass traits was noted in broiler fed guar meal based PROT supplemented diets. Crude protein digestibility, nitrogen retention (Nret) and apparent digestibility coefficient for nitrogen (ADCN) were improved (P<0.05) only with PROT. The improvement in apparent metabolizable energy (AME) and apparent metabolizable energy corrected for nitrogen (AMEn) was noted (P<0.05) with both supplemented enzymes. However, no effect (P>0.05) of enzyme addition was noted on blood glucose, total protein and cholesterol. Improved villus height of duodenum, jejunum and ileum was noted (P<0.05) with the addition of both enzymes. The EAAs digestibility was improved (P<0.05) only with PROT. In conclusion, beta-mannanase and protease supplementation better improved the overall bird performance in low nutrient profile guar meal based diets than non-supplemented diets.

Keywords: novel protease, guar meal, broilers, low protein diets, low metabolizable energy diets, nutrients digestibility

Procedia PDF Downloads 62

Authors: P. S. BEDI* , Neavty Thakur, Balvınder Sıngh

Abstract:

In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3, NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria eg. Bacillus subtilis, Gram negative bacteria eg. E. coli, K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant posseses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Keywords: Girardinia heterophylla, leaves and stem extracts, Antibacterial activity, antifungal activity.

Procedia PDF Downloads 345

Authors: Mohamed E. Khalifa, Adil A. Gobouri

Abstract:

Potential synthesis of a series of 3-amino-4-arylazothiophene derivatives from reaction of 2-cyano-2-phenylthiocarbamoyl acetamide and the appropriate α-halogenated reagents, followed by coupling with different aryl diazonium salts (Japp-Klingemann reaction), and another series of 5-arylazo-thiazol-2-ylcarbamoyl-thiophene derivatives from base-catalyzed intramolecular condensation of 5-arylazo-2-(N-chloroacetyl)amino-thiazole with selected B-keto compounds (Thorpe-Ziegler reaction) was performed. The biological activity of the two series was studied in vitro. Their versatility for pharmaceutical purposes was reported, where they displayed remarkable activities against selected pathogenic microorganisms; Bacillus subtilize, Staphylococcus aureus (Gram positive bacteria), Escherichia coli, Pseudomonas aeruginosa (Gram negative bacteria) and Aspergillus flavus, Candida albicans (fungi) with various degrees related to their chemical structures.

Keywords: thiophene, 2-aminothiazole, compounds, antioxidant, antitumor, antimicrobial

Procedia PDF Downloads 344

Authors: C. Makwana, S. R. Dave

Abstract:

Arsenic is toxic to almost all living cells. Its contamination in natural sources affects the growth of microorganisms. The presence of arsenic is associated with various human disorders also. The attempt of this sort of study provides information regarding the performance of our isolated microorganisms in the presence of Arsenic, which have ample scope for bioremediation. Six isolates were selected from the polluted sample of industrial zone Vatva, Ahmedabad, Gujarat, India, out of which two were Thermophilic organisms. The thermophilic exopolysaccharide (EPS) producing Bacillus was used for microbial enhance oil recovery (MEOR) and in the bio beneficiation. Inorganic arsenic primarily exists in the form of arsenate or arsenite. This arsenic resistance isolate was capable of transforming As +3 to As+5. This isolate would be useful for arsenic remediation standpoint from aquatic systems. The study revealed that the thermophilic microorganism was growing at 55 degree centigrade showed considerable remediation property. The results on the growth and enzyme catalysis would be discussed in response to Arsenic remediation.

Keywords: aquatic systems, thermophilic, exopolysacchride, arsenic

Procedia PDF Downloads 213

Authors: M. A. El-Khateeb

Abstract:

The growth of the olive oil production in Saudi Arabia peculiarly in Al Jouf region in recent years has been accompanied by an increase in the discharge of associated processing waste. Olive mill waste is produced throughout the extraction of oil from the olive fruit using the traditional mill and press process. Deterioration of the environment due to olive mill disposal wastes is a serious problem. When olive mill waste disposed into the soil, it affects soil quality, soil micro flora, and also toxic to plants. The aim of this work is to isolate microorganism (bacterial or fungal strains) from OMW capable of degrading phenols. Olive mill wastewater, olive mill waste and soil (beside oil production mill) contaminated with olive waste were used for isolation of phenol tolerant microorganisms. Four strains (two fungal and two bacterial) were isolated from olive mill waste. The isolated strains were Candida tropicalis and Phanerochaete chrysosporium (fungal strains) and Bacillus sp. and Rhodococcus sp. (bacterial strains). These strains were able to degrade phenols and could be used for bioremediation of olive mill waste.

Keywords: bioremediation, bacteria, fungi, Sakaka

Procedia PDF Downloads 362

Authors: Ghazala Yaqub, Zubi Sadiq, Almas Hamid, Saira Iqbal

Abstract:

This paper is the very first report of three dual action hybrids synthesized by covalent linkage of carbazole based novel antibacterial compounds with efflux pump inhibitors i.e., indole acetic acid/gallic acid. Novel carbazole based antibacterial compounds were prepared first and then these were covalently linked with efflux pump inhibitors which leads to the successful formation of hybrids. All prepared compounds were evaluated for their bacterial cell killing capability against Escherichia coli, Staphylococcus aureus, Pasteurella multocida and Bacillus subtilis. Compound were effective against all tested bacterial strains at different concentrations. But when these compounds were linked with efflux pump inhibitors they showed dramatic enhancement in their bacterial cell killing potential and minimum inhibitory concentration of all hybrids ranges from 7.250 µg/mL to 0.0283 µg/mL.

Keywords: antimicrobial assay, carbazole, dual action hybrids, efflux pump inhibitors

Procedia PDF Downloads 2104

Authors: Satish Babu Rajulapati

Abstract:

The removal of toxic and heavy metal contaminants from wastewater streams and industrial effluents is one of the most important environmental issues being faced world over. In the present study three bacterial cultures tolerating high concentrations of chromium were isolated from the soil and wastewater sample collected from the tanneries located in Warangal, Telangana state. The bacterial species were identified as Bacillus sp., Staphylococcus sp. and pseudomonas sp. Preliminary studies were carried out with the three bacterial species at various operating parameters such as pH and temperature. The results indicate that pseudomonas sp. is the efficient one in the uptake of Cr(VI). Further, detailed investigation of Pseudomonas sp. have been carried out to determine the efficiency of removal of Cr(VI). The various parameters influencing the biosorption of Cr(VI) such as pH, temperature, initial chromium concentration, innoculum size and incubation time have been studied. Response Surface Methodology (RSM) was applied to optimize the removal of Cr(VI). Maximum Cr(VI) removal was found to be 85.72% Cr(VI) atpH 7, temperature 35 °C, initial concentration 67mg/l, inoculums size 9 %(v/v) and time 60 hrs.

Keywords: Staphylococcus sp, chromium, RSM, optimization, Cr(IV)

Procedia PDF Downloads 324