Search results for: resistant starch
937 Antibiotic Potential of Bioactive Compounds from a Marine Streptomyces Isolated from South Pacific Sediments
Authors: Ilaisa Kacivakanadina, Samson Viulu, Brad Carte, Katy Soapi
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Two bioactive compounds namely Vulgamycin (also known as enterocin A) and 5-deoxyenterocin were purified from a marine bacterial strain 1903. Strain 1903 was isolated from marine sediments collected from the Solomon Islands. Morphological features of strain 1903 showed that it belongs to the genus Streptomyces. The two secondary metabolites were extracted using EtOAc and purified by chromatographic methods using EtOAc and hexane solvents. Mass spectrum and NMR data of pure compounds were used to elucidate the chemical structures. In this study, results showed that both compounds were strongly active against Wild Type Staphylococcus aureus (WTSA) (MIC < 1 µg/mL) and in Brine shrimp assays (BSA) (MIC < 1 µg/mL). 5-deoxyenterocin was also active against Rifamycin resistant Staphylococcus aureus (RRSA) (MIC, 250 µg/mL) while vulgamycin showed bioactivity against Methicillin resistant Staphylococcus aureus (MRSA) (MIC 250 µg/mL). To the best of our knowledge, this is the first study that showed the bio-activity of 5-deoxyenterocin. This is also the first time that Vulgamycin has been reported to be active in a BSA. There has not been any mechanism of action studies for these two compounds against pathogens. This warrants further studies on their mechanism of action against microbial pathogens.Keywords: 5-deoxyenterocin, bioactivity, brine shrimp assay (BSA), vulgamycin
Procedia PDF Downloads 179936 Exploring Fluoroquinolone-Resistance Dynamics Using a Distinct in Vitro Fermentation Chicken Caeca Model
Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.
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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model
Procedia PDF Downloads 53935 Discrimination of Bio-Analytes by Using Two-Dimensional Nano Sensor Array
Authors: P. Behera, K. K. Singh, D. K. Saini, M. De
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Implementation of 2D materials in the detection of bio analytes is highly advantageous in the field of sensing because of its high surface to volume ratio. We have designed our sensor array with different cationic two-dimensional MoS₂, where surface modification was achieved by cationic thiol ligands with different functionality. Green fluorescent protein (GFP) was chosen as signal transducers for its biocompatibility and anionic nature, which can bind to the cationic MoS₂ surface easily, followed by fluorescence quenching. The addition of bio-analyte to the sensor can decomplex the cationic MoS₂ and GFP conjugates, followed by the regeneration of GFP fluorescence. The fluorescence response pattern belongs to various analytes collected and transformed to linear discriminant analysis (LDA) for classification. At first, 15 different proteins having wide range of molecular weight and isoelectric points were successfully discriminated at 50 nM with detection limit of 1 nM. The sensor system was also executed in biofluids such as serum, where 10 different proteins at 2.5 μM were well separated. After successful discrimination of protein analytes, the sensor array was implemented for bacteria sensing. Six different bacteria were successfully classified at OD = 0.05 with a detection limit corresponding to OD = 0.005. The optimized sensor array was able to classify uropathogens from non-uropathogens in urine medium. Further, the technique was applied for discrimination of bacteria possessing resistance to different types and amounts of drugs. We found out the mechanism of sensing through optical and electrodynamic studies, which indicates the interaction between bacteria with the sensor system was mainly due to electrostatic force of interactions, but the separation of native bacteria from their drug resistant variant was due to Van der Waals forces. There are two ways bacteria can be detected, i.e., through bacterial cells and lysates. The bacterial lysates contain intracellular information and also safe to analysis as it does not contain live cells. Lysates of different drug resistant bacteria were patterned effectively from the native strain. From unknown sample analysis, we found that discrimination of bacterial cells is more sensitive than that of lysates. But the analyst can prefer bacterial lysates over live cells for safer analysis.Keywords: array-based sensing, drug resistant bacteria, linear discriminant analysis, two-dimensional MoS₂
Procedia PDF Downloads 135934 Affordable and Sustainable Housing Construction: Case Studies
Authors: Tony Rizk
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Recent material advances and cost efficiencies are transforming the housing industry away from traditional lumber and gypsum material to alternate fiberboard material that is workable and resistant to fire, mold, and pest infestation. The use of these materials may add to the initial cost of construction. However, the life cycle (cradle to grave) cost of houses using these construction materials and methods are lower than the life cycle costs using traditional housing construction materials and methods. This paper will present four (4) case studies of sustainable house projects. Each project was designed and constructed using earthen-based, sustainable fiberboard material that is resistant to fire, mold, and infestation and fabricated at a very low material calorific value. These house projects have a living space ranging from 625 sq. ft. for an accessory dwelling unit and up to 3,200 sq. ft. 1-story and 2-story homes. For each case study, we will present the house engineering design and construction method, the initial construction costs, a summary of the life cycle costs, and a comparison to the life cycle cost of traditional housing available in the literature.Keywords: residential housing, sustainable housing, life cycle cost, fire resistance, mold, infestation resistance
Procedia PDF Downloads 119933 The Antimicrobial Activity of Marjoram Essential Oil Against Some Antibiotic Resistant Microbes Isolated from Hospitals
Authors: R. A. Abdel Rahman, A. E. Abdel Wahab, E. A. Goghneimy, H. F. Mohamed, E. M. Salama
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Infectious diseases are a major cause of death worldwide. The treatment of infections continues to be problematic in modern time because of the severe side effects of some drugs and the growing resistance to antimicrobial agents. Hence, the search for newer, safer and more potent antimicrobials is a pressing need. Herbal medicines have received much attention as a source of new antibacterial drugs since they are considered time-tested and comparatively safe both for human use and the environment. In the present study, the antimicrobial activity of marjoram (Origanum majorana L.) essential oil on some gram positive and gram negative reference bacteria, as well as some hospital resistant microbes, was tested. Marjoram oil was extracted and the oil chemical constituents were identified using GC/MS analysis. Staphylococcus aureas ATCC 6923, Pseudomonus auregonosa ATCC 9027, Bacillus subtilis ATCC 6633, E. coli ATCC 8736 and two hospital resistant microbes isolates 16 and 21 were used. The two isolates were identified by biochemical tests and 16s rRNA as proteus spp. and Enterococcus facielus. The effect of different concentrations of essential oils on bacterial growth was tested using agar disk diffusion assay method to determine the minimum inhibitory concentrations and using micro dilution method to determine the minimum bactericidal concentrations. Marjoram oil was found to be effective against both reference and hospital resistance strains. Hospital strains were more resistant to marjoram oil than reference strains. P. auregonosa growth was completely inhibited at a low concentration of oil (4µl/ml). The other reference strains showed sensitivity to marjoram oil at concentrations ranged from 5 to 7µl/ml. The two hospital strains showed sensitivity at media containing 10 and 15µl/ml oil. The major components of oil were terpineol, cis-beta (23.5%), 1,6 – octadien –3-ol,3,7-dimethyl, 2 aminobenzoate (10.9%), alpha terpieol (8.6%) and linalool (6.3%). Scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis were used to determine the difference between treated and untreated hospital strains. SEM results showed that treated cells were smaller in size than control cells. TEM data showed that cell lysis has occurred to treated cells. Treated cells have ruptured cell wall and appeared empty of cytoplasm compared to control cells which shown to be intact with normal volume of cytoplasm. The results indicated that marjoram oil has a positive antimicrobial effect on hospital resistance microbes. Natural crude extracts can be perfect resources for new antimicrobial drugs.Keywords: antimicrobial activity, essential oil, hospital resistance microbes, marjoram
Procedia PDF Downloads 441932 Isolation and Characterization of Anti-melanoma (Skin Cancer) Compounds from Corchorus olitorius .L
Authors: Peramachi Sathiyamoorthy, Jacop Gopas, Avi Golan Goldhirsh
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Corchorus olitorius is a leafy vegetable and an industrial crop. The herb has antioxidant, anti inflammatory, and anti-cancer properties. To assay the pharmaceutical properties, aqueous extracts of leaves and seeds from C. olitorius were tested against drug resistant melanoma cell line. The test showed LC50 of the extract was 0.08µg/ml. Aqueous seed extract exhibited higher melanoma inhibiting activity than leaf extract. Dialysis of seed extract showed that the active compound is less than 12 KDa. The compound with <3 KDa MW separated by microconcentration of seed extract showed 70.5 % inhibition of melanoma cell growth. Among the two fractions obtained by Gel filtration with G10 column, the first fraction at 1:2000 dilutions exhibited 100% inhibition of melanoma growth. The compound with Rf value 0.86 (MA4) isolated by TLC separation showed about 98% cytotoxicity against melanoma at 1: 1000 dilutions. Furthermore, HPLC separation of MA4 compound with Superdex 75 column resulted in 4 compounds. Out of 4, one compound showed melanoma inhibition. The active compound is identified by reagent methods as Strophanthidin. Further toxicological and clinical studies will lead to the development of a potential drug to treat drug resistant melanoma.Keywords: corchorus olitorius, melanoma, drug development, strophanthidin
Procedia PDF Downloads 127931 Spatial Analysis of Survival Pattern and Treatment Outcomes of Multi-Drug Resistant Tuberculosis (MDR-TB) Patients in Lagos, Nigeria
Authors: Akinsola Oluwatosin, Udofia Samuel, Odofin Mayowa
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The study is aimed at assessing the Geographic Information System (GIS)-based spatial analysis of Survival Pattern and Treatment Outcomes of Multi-Drug Resistant Tuberculosis (MDR-TB) cases for Lagos, Nigeria, with an objective to inform priority areas for public health planning and resource allocation. Multi-drug resistant tuberculosis (MDR-TB) develops due to problems such as irregular drug supply, poor drug quality, inappropriate prescription, and poor adherence to treatment. The shapefile(s) for this study were already georeferenced to Minna datum. The patient’s information was acquired on MS Excel and later converted to . CSV file for easy processing to ArcMap from various hospitals. To superimpose the patient’s information the spatial data, the addresses was geocoded to generate the longitude and latitude of the patients. The database was used for the SQL query to the various pattern of the treatment. To show the pattern of disease spread, spatial autocorrelation analysis was used. The result was displayed in a graphical format showing the areas of dispersing, random and clustered of patients in the study area. Hot and cold spot analysis was analyzed to show high-density areas. The distance between these patients and the closest health facility was examined using the buffer analysis. The result shows that 22% of the points were successfully matched, while 15% were tied. However, the result table shows that a greater percentage of it was unmatched; this is evident in the fact that most of the streets within the State are unnamed, and then again, most of the patients are likely to supply the wrong addresses. MDR-TB patients of all age groups are concentrated within Lagos-Mainland, Shomolu, Mushin, Surulere, Oshodi-Isolo, and Ifelodun LGAs. MDR-TB patients between the age group of 30-47 years had the highest number and were identified to be about 184 in number. The outcome of patients on ART treatment revealed that a high number of patients (300) were not ART treatment while a paltry 45 patients were on ART treatment. The result shows the Z-score of the distribution is greater than 1 (>2.58), which means that the distribution is highly clustered at a significance level of 0.01.Keywords: tuberculosis, patients, treatment, GIS, MDR-TB
Procedia PDF Downloads 146930 A Significant Clinical Role for the Capitalbio™ DNA Microarray in the Diagnosis of Multidrug-Resistant Tuberculosis in Patients with Tuberculous Spondylitis Simultaneous with Pulmonary Tuberculosis in High Prevalence Settings in China
Authors: Wenjie Wu, Peng Cheng, Zehua Zhang, Fei Luo, Feng Wu, Min Zhong, Jianzhong Xu
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Background: There has been limited research into the therapeutic efficacy of rapid diagnosis of spinal tuberculosis complicated with pulmonary tuberculosis. We attempted to discover whether the utilization of a DNA microarray assay to detect multidrug-resistant spinal tuberculosis complicated with pulmonary tuberculosis can improve clinical outcomes. Methods: A prospective study was conducted from February 2006 to September 2015. One hundred and forty-three consecutive culture–confirmed, clinically and imaging diagnosed MDR-TB patients with spinal tuberculosis complicated by pulmonary tuberculosis were enrolled into the study. The initial time to treatment for MDR-TB, the method of infection control, radiological indicators of spinal tubercular infectious foci, culture conversion, and adverse drug reactions were compared with the standard culture methods. Results: Of the total of 143 MDR-TB patients, 68 (47.6%) were diagnosed by conventional culture methods and 75 (52.4%) following the implementation of detection using the DNA microarray. Patients in the microarray group began rational use of the second-line drugs schedule more speedily than sufferers in the culture group (17.3 vs. 74.1 days). Among patients were admitted to a general tuberculosis ward, those from the microarray group spent less time in the ward than those from the culture group (7.8 vs. 49.2 days). In those patients with six months follow-up (n=134), patients in the microarray group had a higher rate of sputum negativity conversion at six months (89% vs. 73%). In the microarray group, the rate of drug adverse reactions was significantly lower (22.2% vs. 67.7%). At the same time, they had a more obvious reduction of the area with spinal tuberculous lesions in radiological examinations (77% vs. 108%). Conclusions: The application of the CapitalBio™ DNA Microarray assay caused noteworthy clinical advances including an earlier time to begin MDR-TB treatment, increased sputum culture conversion, improved infection control measures and better radiographical resultsKeywords: tuberculosis, multidrug-resistant, tuberculous spondylitis, DNA microarray, clinical outcomes
Procedia PDF Downloads 280929 Growth and Some Physiological Properties of Three Selected Species of Bifidobacteria in Admixture of Soy Milk and Goat Milk
Authors: Ahmed Zahran
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Bifidobacterium breve ATCC 15700, Bifidobacterium adolescents ATCC 15704 and Bifidobacterium longum ATCC 15707 were tested for their growth, acid production, bile tolerance, antibiotic resistance and adherence to columnar epithelial cells of the small intestine of goat. The growth of all studied species was determined in the MRSL medium. B.longum 15707 was the most active species in comparison with the other two species; it was also more resistant to bile acids. The adhesion of the studied species to the columnar epithelial cells was studied. All the studied species showed some degree of adhesion; however, B.longum adhered more than the other two species. This species was resistant to four types of antibiotics and was sensitive to chloramphenicol 30 µg. The activity of Bifidobacterium species in soymilk was evaluated by measuring the development of titratalle acidity. B.longum 15707 was the most active species in terms of growth and activity of soymilk. So, soymilk containing bifidobacteria could be added to goat milk to produce acceptable functional soy yogurt, using the ratio of (1:4) soy milk to goat milk. This product could be of unique health benefits, especially in the case of high cholesterol levels and replenishment of intestinal flora after antibiotic therapy.Keywords: bifidobacteria physiological properties, soy milk, goat milk, attachment epithelial cells, columnar tissues, probiotic food
Procedia PDF Downloads 75928 Anti-tuberculosis, Resistance Modulatory, Anti-pulmonary Fibrosis and Anti-silicosis Effects of Crinum Asiaticum Bulbs and Its Active Metabolite, Betulin
Authors: Theophilus Asante, Comfort Nyarko, Daniel Antwi
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Drug-resistant tuberculosis, together with the associated comorbidities like pulmonary fibrosis and silicosis, has been one of the most serious global public health threats that requires immediate action to curb or mitigate it. This prolongs hospital stays, increases the cost of medication, and increases the death toll recorded annually. Crinum asiaticum bulb (CAE) and betulin (BET) are known for their biological and pharmacological effects. Pharmacological effects reported on CAE include antimicrobial, anti-inflammatory, anti-pyretic, anti-analgesic, and anti-cancer effects. Betulin has exhibited a multitude of powerful pharmacological properties ranging from antitumor, anti-inflammatory, anti-parasitic, anti-microbial, and anti-viral activities. This work sought to investigate the anti-tuberculosis and resistant modulatory effects and also assess their effects on mitigating pulmonary fibrosis and silicosis. In the anti-tuberculosis and resistant modulatory effects, both CAE and BET showed strong antimicrobial activities (31.25 ≤ MIC ≤ 500) µg/ml against the studied microorganisms and also produced significant anti-efflux pump and biofilm inhibitory effects (ρ < 0.0001) as well as exhibiting resistance modulatory and synergistic effects when combined with standard antibiotics. Crinum asiaticum bulbs extract and betulin were shown to possess anti-pulmonary fibrosis effects. There was an increased survival rate in the CAE and BET treatment groups compared to the BLM-induced group. There was a marked decrease in the levels of hydroxyproline and collagen I and III in the CAE and BET treatment groups compared to the BLM-treated group. The treatment groups of CAE and BET significantly downregulated the levels of pro-fibrotic and pro-inflammatory cytokine concentrations such as TGF-β1, MMP9, IL-6, IL-1β and TNF-alpha compared to an increase in the BLM-treated groups. The histological findings of the lungs suggested the curative effects of CAE and BET following BLM-induced pulmonary fibrosis in mice. The study showed improved lung functions with a wide focal area of viable alveolar spaces and few collagen fibers deposition on the lungs of the treatment groups. In the anti-silicosis and pulmonoprotective effects of CAE and BET, the levels of NF-κB, TNF-α, IL-1β, IL-6 and hydroxyproline, collagen types I and III were significantly reduced by CAE and BET (ρ < 0.0001). Both CAE and BET significantly (ρ < 0.0001) inhibited the levels of hydroxyproline, collagen I and III when compared with the negative control group. On BALF biomarkers such as macrophages, lymphocytes, monocytes, and neutrophils, CAE and BET were able to reduce their levels significantly (ρ < 0.0001). The CAE and BET were examined for anti-oxidant activity and shown to raise the levels of catalase (CAT) and superoxide dismutase (SOD) while lowering the level of malondialdehyde (MDA). There was an improvement in lung function when lung tissues were examined histologically. Crinum asiaticum bulbs extract and betulin were discovered to exhibit anti-tubercular and resistance-modulatory properties, as well as the capacity to minimize TB comorbidities such as pulmonary fibrosis and silicosis. In addition, CAE and BET may act as protective mechanisms, facilitating the preservation of the lung's physiological integrity. The outcomes of this study might pave the way for the development of leads for producing single medications for the management of drug-resistant tuberculosis and its accompanying comorbidities.Keywords: fibrosis, crinum, tuberculosis, antiinflammation, drug resistant
Procedia PDF Downloads 79927 ROCK Signaling and Radio Resistance: The Association and the Effect
Authors: P. Annapurna, Cecil Ross, Sudhir Krishna, Sweta Srivastava
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Irradiation plays a pivotal role in cervical cancer treatment, however some tumors exhibit resistance to therapy while some exhibit relapse, due to better repair and enhanced resistance mechanisms operational in their cells. The present study aims to understand the signaling mechanism operational in resistance phenotype and in the present study we report the role of Rho GTPase associated protein kinase (ROCK) signaling in cervical carcinoma radio-resistance. ROCK signaling has been implicated in several tumor progressions and is important for DNA repair. Irradiation of spheroid cultures of SiHa cervical carcinoma derived cell line at 6Gy resulted in generation of resistant cells in vitro which had better clonogenic abilities and formed larger and more colonies, in soft agar colony formation assay, as compared to the non-irradiated cells. These cells also exhibited an enhanced motility phenotype. Cell cycle profiling showed the cells to be blocked in G2M phase with enhanced pCDC2 levels indicating onset of possible DNA repair mechanism. Notably, 3 days post-irradiation, irradiated cells showed increased ROCK2 translocation to the nucleus with enhanced protein expression as compared to the non-irradiated cells. Radio-sensitization of the resistant cells was enhanced using Y27632, an inhibitor to ROCK signaling. The treatment of resistant cells with Y27632 resulted in increased cell death upon further irradiation. This observation has been confirmed using inhibitory antibodies to ROCK1/2. Result show that both ROCK1/2 have a functional contribution in radiation resistance of cervical cancer cells derived from cell lines. Interestingly enrichment of stem like cells (Hoechst negative cells) was also observed upon irradiation and these cells were markedly sensitive to Y27632 treatment. Our results thus suggest the role of ROCK signaling in radio-resistance in cervical carcinoma. Further studies with human biopsies, mice models and mechanistic of ROCK signaling in the context of radio-resistance will clarify the role of this molecule further and allow for therapeutics development.Keywords: cervical carcinoma, radio-resistance, ROCK signaling, cancer treatment
Procedia PDF Downloads 316926 The Biology of Persister Cells and Antibiotic Resistance
Authors: Zikora K. G. Anyaegbunam, Annabel A. Nnawuihe, Ngozi J. Anyaegbunam, Emmanuel A. Eze
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The discovery and production of new antibiotics is unavoidable in the fight against drug-resistant bacteria. However, this is only part of the problem; we have never really had medications that could completely eradicate an infection. All pathogens create a limited number of dormant persister cells that are resistant to antibiotic treatment. When the concentration of antibiotics decreases, surviving persisters repopulate the population, resulting in a recurrent chronic infection. Bacterial populations have an alternative survival strategy to withstand harsh conditions or antibiotic exposure, in addition to the well-known methods of antibiotic resistance and biofilm formation. Persister cells are a limited subset of transiently antibiotic-tolerant phenotypic variations capable of surviving high-dose antibiotic therapy. Persisters that flip back to a normal phenotype can restart growth when antibiotic pressure drops, assuring the bacterial population's survival. Persister cells have been found in every major pathogen, and they play a role in antibiotic tolerance in biofilms as well as the recalcitrance of chronic infections. Persister cells has been implicated to play a role in the establishment of antibiotic resistance, according to growing research. Thusthe need to basically elucidate the biology of persisters and how they are linked to antibiotic resistance, and as well it's link to diseases.Keywords: persister cells, phenotypic variations, repopulation, mobile genetic transfers, antibiotic resistance
Procedia PDF Downloads 200925 The Effect of Durability and Pathogen Strains on the Wheat Induced Resistance against Zymoseptoria tritici as a Response to Paenibacillus sp. Strain B2
Authors: E. Samain, T. Aussenac, D. van Tuinen, S. Selim
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Plant growth promoting rhizobacteria are known as potential biofertilizers and plant resistance inducers. The present work aims to study the durability of the resistance induced as a response to wheat seeds inoculation with PB2 and its influence by Z. tritici strains. The internal and external roots colonization have been determined in vitro, seven days post inoculation, by measuring the colony forming unit (CFU). In planta experimentations were done under controlled conditions included four wheat cultivars with different levels of resistance against Septoria Leaf Blotch (SLB) and four Z. tritici strains with high aggressiveness and resistance levels to fungicides. Plantlets were inoculated with PB2 at sowing and infected with Z. tritici at 3 leaves or tillering growth stages. The infection level with SLB was evaluated at 17 days post inoculation using real-time quantitative polymerase chain reaction (PCR). Results showed that PB2 has a high potential of wheat root external colonization (> 10⁶ CFU/g of root). However, the internal colonization seems to be cultivar dependent. Indeed, PB2 has not been observed as endophytic for one cultivar but has a high level of internal colonization with more than 104 CFU/g of root concerning the three others. Two wheat cultivars (susceptible and moderated resistant) were used to investigate PB2-induced resistance (PB2-IR). After the first infection with Z. tritici, results showed that PB2-IR has conferred a high protection efficiency (40-90%) against SLB in the two tested cultivars. Whereas the PB2-IR was effective against all tested strains with the moderate resistant cultivar, it was higher with the susceptible cultivar (> 64%) but against three of the four tested strains. Concerning the durability of the PB2-IR, after the second infection timing, it has been observed a significant decrease (10-59%) depending strains in the moderate resistant cultivar. Contrarily, the susceptible cultivar showed a stable and high protection level (76-84%) but against three of the four tested strains and interestingly, the strain that overcame PB2-IR was not the same as that of the first infection timing. To conclude, PB2 induces a high and durable resistance against Z. tritici. The PB2-IR is pathogen strain, plant growth stage and genotype dependent. These results may explain the loss of the induced resistance effectiveness under field conditions.Keywords: induced resistance, Paenibacillus sp. strain B2, wheat genotypes, Zymoseptoria tritici
Procedia PDF Downloads 142924 Investigation of Carbapenem-Resistant Genes in Acinetobacter spp. Isolated from Patients at Tertiary Health Care Center, Northeastern Thailand
Authors: S. J. Sirima, C. Thirawan, R.Puntharikorn, K. Ungsumalin, J. Kaemwich
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Acinetobacter spp. is a gram negative bacterium causing the high incidence of multi-drug resistance in patients admitted to an intensive care unit. A hundred isolates of Imipenem-resistant Acinetobacter spp. isolated from patients admitted at tertiary health care center, Northeastern region, Ubon Ratchathani, Thailand, were subjected to modified Hodge test and combined disc test in order to evaluate the production of carbapenemases. The results revealed that about 35% of isolates were found to be carbapenemases producers. In addition, multiplex polymerase chain reactions were performed to detect blaOXA-like genes. It showed that 92% of isolates possess blaOXA-51-like and blaOXA-23-like genes. However, blaOXA-58-like gene was detected in only 8 isolates. No detection of blaOXA-24-like gene was observed in all isolates. In conclusion, an ability to produce carbepenemases would be an important mechanism of multi-drug resistance among clinical isolates of Acinetobacter spp. at tertiary health care center, Northeastern region, Ubon Ratchathani, Thailand. Furthermore, it was likely that the class D carbapenemases genes, blaOXA-51-like and blaOXA-23-like, might contribute to imipenem-resistance exhibiting among isolates.Keywords: Acinetobacter spp., blaOXA-like genes, carbapenemases, tertiary health care center
Procedia PDF Downloads 376923 Characteristics of Sorghum (Sorghum bicolor L. Moench) Flour on the Soaking Time of Peeled Grains and Particle Size Treatment
Authors: Sri Satya Antarlina, Elok Zubaidah, Teti Istiana, Harijono
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Sorghum bicolor (Sorghum bicolor L. Moench) has the potential as a flour for gluten-free food products. Sorghum flour production needs grain soaking treatment. Soaking can reduce the tannin content which is an anti-nutrient, so it can increase the protein digestibility. Fine particle size decreases the yield of flour, so it is necessary to study various particle sizes to increase the yield. This study aims to determine the characteristics of sorghum flour in the treatment of soaking peeled grain and particle size. The material of white sorghum varieties KD-4 from farmers in East Java, Indonesia. Factorial randomized factorial design (two factors), repeated three times, factor I were the time of grain soaking (five levels) that were 0, 12, 24, 36, and 48 hours, factor II was the size of the starch particles sifted with a fineness level of 40, 60, 80, and 100 mesh. The method of making sorghum flour is grain peeling, soaking peeled grain, drying using the oven at 60ᵒC, milling, and sieving. Physico-chemical analysis of sorghum flour. The results show that there is an interaction between soaking time of grain with the size of sorghum flour particles. Interaction in yield of flour, L* color (brightness level), whiteness index, paste properties, amylose content, protein content, bulk density, and protein digestibility. The method of making sorghum flour through the soaking of peeled grain and the difference in particle size has an important role in producing the physicochemical properties of the specific flour. Based on the characteristics of sorghum flour produced, it is determined the method of making sorghum flour through sorghum grain soaking for 24 hours, the particle size of flour 80 mesh. The sorghum flour with characteristic were 24.88% yield of flour, 88.60 color L* (brightness level), 69.95 whiteness index, 3615 Cp viscosity, 584.10 g/l of bulk density, 24.27% db protein digestibility, 90.02% db starch content, 23.4% db amylose content, 67.45% db amylopectin content, 0.22% db crude fiber content, 0.037% db tannin content, 5.30% db protein content, ash content 0.18% db, carbohydrate content 92.88 % db, and 1.94% db fat content. The sorghum flour is recommended for cookies products.Keywords: characteristic, sorghum (Sorghum bicolor L. Moench) flour, grain soaking, particle size, physicochemical properties
Procedia PDF Downloads 153922 Tuberculosis Outpatient Treatment in the Context of Reformation of the Health Care System
Authors: Danylo Brindak, Viktor Liashko, Olexander Chepurniy
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Despite considerable experience in implementation of the best international approaches and services within response to epidemy of multi-drug resistant tuberculosis, the results of situation analysis indicate the presence of faults in this area. In 2014, Ukraine (for the first time) was included in the world’s five countries with the highest level of drug-resistant tuberculosis. The effectiveness of its treatment constitutes only 35% in the country. In this context, the increase in allocation of funds to control the epidemic of multidrug-resistant tuberculosis does not produce perceptible positive results. During 2001-2016, only the Global Fund to fight AIDS, Tuberculosis, and Malaria allocated to Ukraine more than USD 521,3 million for programs of tuberculosis and HIV/AIDS control. However, current conditions in post-Semashko system create little motivation for rational use of resources or cost control at inpatient TB facilities. There is no motivation to reduce overdue hospitalization and to target resources to priority sectors of modern tuberculosis control, including a model of care focused on the patient. In the presence of a line-item budget at medical institutions, based on the input factors as the ratios of beds and staff, there is a passive disposal of budgetary funds by health care institutions and their employees who have no motivation to improve quality and efficiency of service provision. Outpatient treatment of tuberculosis is being implemented in Ukraine since 2011 and has many risks, namely creation of parallel systems, low consistency through dependence on funding for the project, reduced the role of the family doctor, the fragmentation of financing, etc. In terms of reforming approaches to health system financing, which began in Ukraine in late 2016, NGO Infection Control in Ukraine conducted piloting of a new, motivating method of remuneration of employees in primary health care. The innovative aspect of this funding mechanism is cost according to results of treatment. The existing method of payment on the basis of the standard per inhabitant (per capita ratio) was added with motivating costs according to results of work. The effectiveness of such treatment of TB patients at the outpatient stage is 90%, while in whole on the basis of a current system the effectiveness of treatment of newly diagnosed pulmonary TB with positive swab is around 60% in the country. Even though Ukraine has 5.24 TB beds per 10 000 citizens. Implemented pilot model of ambulatory treatment will be used for the creation of costs system according to results of activities, the integration of TB and primary health and social services and their focus on achieving results, the reduction of inpatient treatment of tuberculosis.Keywords: health care reform, multi-drug resistant tuberculosis, outpatient treatment efficiency, tuberculosis
Procedia PDF Downloads 145921 Purification and Pre-Crystallization of Recombinant PhoR Cytoplasmic Domain Protein from Mycobacterium Tuberculosis H37Rv
Authors: Oktira Roka Aji, Maelita R. Moeis, Ihsanawati, Ernawati A. Giri-Rachman
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Globally, tuberculosis (TB) remains a leading cause of death. The emergence of multidrug-resistant strains and extensively drug-resistant strains have become a major public concern. One of the potential candidates for drug target is the cytoplasmic domain of PhoR Histidine Kinase, a part of the Two Component System (TCS) PhoR-PhoP in Mycobacterium tuberculosis (Mtb). TCS PhoR-PhoP relay extracellular signal to control the expression of 114 virulent associated genes in Mtb. The 3D structure of PhoR cytoplasmic domain is needed to screen novel drugs using structure based drug discovery. The PhoR cytoplasmic domain from Mtb H37Rv was overexpressed in E. coli BL21(DE3), then purified using IMAC Ni-NTA Agarose his-tag affinity column and DEAE-ion exchange column chromatography. The molecular weight of the purified protein was estimated to be 37 kDa after SDS-PAGE analysis. This sample was used for pre-crystallization screening by applying sitting drop vapor diffusion method using Natrix (HR2-116) 48 solutions crystal screen kit at 25ºC. Needle-like crystals were observed after the seventh day of incubation in test solution No.47 (0.1 M KCl, 0.01 M MgCl2.6H2O, 0.05 M Tris-Cl pH 8.5, 30% v/v PEG 4000). Further testing is required for confirming the crystal.Keywords: tuberculosis, two component system, histidine kinase, needle-like crystals
Procedia PDF Downloads 428920 COX-2 Inhibitor NS398 Counteracts Chemoresistance to Temozolomide in T98G Glioblastoma Cell Line
Authors: Francesca Lombardi, Francesca Rosaria Augello, Benedetta Cinque, Maria Grazia Cifone, Paola Palumbo
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Glioblastoma multiforme (GBM) is a high-grade primary brain tumor refractory to current forms of treatment. The survival benefits of patients with GBM remain unsatisfactory due to the intrinsic or acquired resistance to temozolomide (TMZ), an alkylating agent, used as the first-line chemotherapeutic drug to treat GBM patients. Its cytotoxic effect is visualized by the induction of O6-methylguanine (O6MeG) within DNA. Cyclooxygenase-2 (COX-2), an inflammation-associated enzyme, has been implicated in tumorigenesis and progression of GBM, its inhibition shows anticancer activities. In the present study, it was verified if the combination of a COX-2 selective inhibitor, NS398, with TMZ could counteract the TMZ resistance. In particular, the effect of NS398 mixed with TMZ was investigated in the GBM TMZ-resistant cell line, T98G. Cells were pretreated with NS398 (100µM, 24 hours) and then exposed to TMZ alone (200µM), NS398 alone, or both for 72 hours, after which cell growth rate and cycle phases, as well as apoptosis level, were evaluated. Coadministration of NS398 and TMZ caused a significant decrease in cell growth and a progressive increase of dead cells detected by trypan blue staining. Moreover, a significant level of apoptotic cell percentage and alteration of cell cycle phases were observed in T98G treated with TMZ-NS398 combination when compared to untreated cells or TMZ-treated cells. TMZ-resistant tumors, as GBM, express elevated levels of DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT). The mixture drastically reduced MGMT expression in the TMZ-resistant cell line T98G, known to express high levels of MGMT basically. Moreover, while TMZ alone did not influence the COX-2 protein expression, the combination successfully reduced it. In conclusion, these results demonstrated that NS398 enhanced the efficacy of TMZ through cell number reduction, apoptosis induction, and decreased MGMT levels, suggesting the ability of drug combination to reduce the chemoresistance. This drug combination deserves attention and could be considered as a promising therapeutic strategy for GBM patients.Keywords: COX-2, COX-2 inhibitor, glioblastoma, NS398, T98G, temozolomide
Procedia PDF Downloads 143919 Isolation and Molecular Characterization of Lytic Bacteriophage against Carbapenem Resistant Klebsiella pneumoniae
Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla
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Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,
Procedia PDF Downloads 186918 Influence of Laser Treatment on the Growth of Sprouts of Different Wheat Varieties
Authors: N. Bakradze, T. Dumbadze, N. Gagelidze, L. Amiranashvili, A. D. L. Batako
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Cereals are considered as a strategic product in human life and it demand is increasing with the growth of world population. There is always shortage of cereals in various areas of the globe. For example, Georgia own production meets only 15-20% of the demand for grain, despite the fact that the country is considered one of the main centers of wheat origin. In Georgia, there are 14 types of wheat and more than 150 subspecies, and 40 subspecies of common wheat. Increasing wheat production is important for the country. One of the ways to solve the problem is to develop and implement new, environmentally and economically acceptable technologies. Such technologies include pre-sowing treatment of seed with a laser and associative nitrogen-fixing of the Azospirillum brasilensse bacteria. In the region there are Dika and Lomtagora which are among the most common in Georgia. Dika is a frost-resistant wheat, with a high ability to adapt to the environment, resistant to falling and it is sown in highlands. Dicka excellent properties are due to its strong immunity to fungal diseases; Dicka grains are rich in protein and lysine. Lomtagora 126 differs with its winter and drought resistance, and, it has a great ability to germinate. Lomtagora is characterized by a strong root system and a high budding capacity. It is an early variety, fall-resistant, easy to thresh and suitable for mechanized harvesting with large and red grains. The plant is moderately resistant to fungal diseases. This paper presents some preliminary experimental results where, a continuous CO2 laser at a power of 25-40 W/cm2 was used to radiate grains at a flow rate of 10-15 cm/sec. The treatment was carried out on grains of the Triticum aestivum L. var. of Lutescens (local variety name - Lomtagora 126), and Triticum carthlicum Nevski (local variety name - Dika). Here the grains were treated with Azospirillum brasilensse isolate (108-109 CFU / ml), which was isolated from the rhizosphere of wheat. It was observed that the germination of the wheat was not significantly influenced by either laser or bacteria treatment. In the case of the variety Lomtagora 126, when irradiated at an angle of 90°, it slightly improved the growth within 38 days of sawing, and in the case of irradiation at an angle of 90°+1, by 23%. The treatment of seeds with Azospirillum brazilense in both irradiated and non-irradiated variants led to an improvement in the growth of ssprouts. However, in the case of treatment with azospiril alone - by 22%, and with joint treatment of seeds with azospiril and irradiation - by 29%. In the case of the Dika wheat, the irradiation only led to an increase in growth by 8-9%, and the combine treatment of seeds with azospiril and irradiation - by 10-15%, in comparison with the control. Thus, the combine treatment of wheat of different varieties provided the best effect on the growth. Acknowledgment: This work was supported by Shota Rustaveli National Science Foundation of Georgia (SRNSFG) (Grant number CARYS 19-573)Keywords: laser treatment, Azospirillum brasilensse, seeds, wheat varieties, Lomtagora, Dika
Procedia PDF Downloads 140917 Understanding the Diversity of Antimicrobial Resistance among Wild Animals, Livestock and Associated Environment in a Rural Ecosystem in Sri Lanka
Authors: B. M. Y. I. Basnayake, G. G. T. Nisansala, P. I. J. B. Wijewickrama, U. S. Weerathunga, K. W. M. Y. D. Gunasekara, N. K. Jayasekera, A. W. Kalupahana, R. S. Kalupahana, A. Silva- Fletcher, K. S. A. Kottawatta
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Antimicrobial resistance (AMR) has attracted significant attention worldwide as an emerging threat to public health. Understanding the role of livestock and wildlife with the shared environment in the maintenance and transmission of AMR is of utmost importance due to its interactions with humans for combating the issue in one health approach. This study aims to investigate the extent of AMR distribution among wild animals, livestock, and environment cohabiting in a rural ecosystem in Sri Lanka: Hambegamuwa. One square km area at Hambegamuwa was mapped using GPS as the sampling area. The study was conducted for a period of five months from November 2020. Voided fecal samples were collected from 130 wild animals, 123 livestock: buffalo, cattle, chicken, and turkey, with 36 soil and 30 water samples associated with livestock and wildlife. From the samples, Escherichia coli (E. coli) was isolated, and their AMR profiles were investigated for 12 antimicrobials using the disk diffusion method following the CLSI standard. Seventy percent (91/130) of wild animals, 93% (115/123) of livestock, 89% (32/36) of soil, and 63% (19/30) of water samples were positive for E. coli. Maximum of two E. coli from each sample to a total of 467 were tested for the sensitivity of which 157, 208, 62, and 40 were from wild animals, livestock, soil, and water, respectively. The highest resistance in E. coli from livestock (13.9%) and wild animals (13.3%) was for ampicillin, followed by streptomycin. Apart from that, E. coli from livestock and wild animals revealed resistance mainly against tetracycline, cefotaxime, trimethoprim/ sulfamethoxazole, and nalidixic acid at levels less than 10%. Ten cefotaxime resistant E. coli were reported from wild animals, including four elephants, two land monitors, a pigeon, a spotted dove, and a monkey which was a significant finding. E. coli from soil samples reflected resistance primarily against ampicillin, streptomycin, and tetracycline at levels less than in livestock/wildlife. Two water samples had cefotaxime resistant E. coli as the only resistant isolates out of 30 water samples tested. Of the total E. coli isolates, 6.4% (30/467) was multi-drug resistant (MDR) which included 18, 9, and 3 isolates from livestock, wild animals, and soil, respectively. Among 18 livestock MDRs, the highest (13/ 18) was from poultry. Nine wild animal MDRs were from spotted dove, pigeon, land monitor, and elephant. Based on CLSI standard criteria, 60 E. coli isolates, of which 40, 16, and 4 from livestock, wild animal, and environment, respectively, were screened for Extended Spectrum β-Lactamase (ESBL) producers. Despite being a rural ecosystem, AMR and MDR are prevalent even at low levels. E. coli from livestock, wild animals, and the environment reflected a similar spectrum of AMR where ampicillin, streptomycin, tetracycline, and cefotaxime being the predominant antimicrobials of resistance. Wild animals may have acquired AMR via direct contact with livestock or via the environment, as antimicrobials are rarely used in wild animals. A source attribution study including the effects of the natural environment to study AMR can be proposed as this less contaminated rural ecosystem alarms the presence of AMR.Keywords: AMR, Escherichia coli, livestock, wildlife
Procedia PDF Downloads 205916 Nutritional Value Determination of Different Varieties of Oats and Barley Using Near-Infrared Spectroscopy Method for the Horses Nutrition
Authors: V. Viliene, V. Sasyte, A. Raceviciute-Stupeliene, R. Gruzauskas
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In horse nutrition, the most suitable cereal for their rations composition could be defined as oats and barley. Oats have high nutritive value because it provides more protein, fiber, iron and zinc than other whole grains, has good taste, and an activity of stimulating metabolic changes in the body. Another cereal – barley is very similar to oats as a feed except for some characteristics that affect how it is used; however, barley is lower in fiber than oats and is classified as a "heavy" feed. The value of oats and barley grain, first of all is dependent on its composition. Near-infrared spectroscopy (NIRS) has long been considered and used as a significant method in component and quality analysis and as an emerging technology for authenticity applications for cereal quality control. This paper presents the chemical and amino acid composition of different varieties of barley and oats, also digestible energy of different cereals for horses. Ten different spring barley (n = 5) and oats (n = 5) varieties, grown in one location in Lithuania, were assayed for their chemical composition (dry matter, crude protein, crude fat, crude ash, crude fiber, starch) and amino acids content, digestible amino acids and amino acids digestibility. Also, the grains digestible energy for horses was calculated. The oats and barley samples reflectance spectra were measured by means of NIRS using Foss-Tecator DS2500 equipment. The chemical components: fat, crude protein, starch and fiber differed statistically (P<0.05) between the oats and barley varieties. The highest total amino acid content between oats was determined in variety Flamingsprofi (4.56 g/kg) and the lowest – variety Circle (3.57 g/kg), and between barley - respectively in varieties Publican (3.50 g/kg) and Sebastian (3.11 g/kg). The different varieties of oats digestible amino acid content varied from 3.11 g/kg to 4.07 g/kg; barley different varieties varied from 2.59 g/kg to 2.94 g/kg. The average amino acids digestibility of oats varied from 74.4% (Liz) to 95.6% (Fen) and in barley - from 75.8 % (Tre) to 89.6% (Fen). The amount of digestible energy in the analyzed varieties of oats and barley was an average compound 13.74 MJ/kg DM and 14.85 MJ/kg DM, respectively. An analysis of the results showed that different varieties of oats compared with barley are preferable for horse nutrition according to the crude fat, crude fiber, ash and separate amino acids content, but the analyzed barley varieties dominated the higher amounts of crude protein, the digestible Liz amount and higher DE content, and thus, could be recommended for making feed formulation for horses combining oats and barley, taking into account the chemical composition of using cereal varieties.Keywords: barley, digestive energy, horses, nutritional value, oats
Procedia PDF Downloads 201915 Remote Assessment and Change Detection of GreenLAI of Cotton Crop Using Different Vegetation Indices
Authors: Ganesh B. Shinde, Vijaya B. Musande
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Cotton crop identification based on the timely information has significant advantage to the different implications of food, economic and environment. Due to the significant advantages, the accurate detection of cotton crop regions using supervised learning procedure is challenging problem in remote sensing. Here, classifiers on the direct image are played a major role but the results are not much satisfactorily. In order to further improve the effectiveness, variety of vegetation indices are proposed in the literature. But, recently, the major challenge is to find the better vegetation indices for the cotton crop identification through the proposed methodology. Accordingly, fuzzy c-means clustering is combined with neural network algorithm, trained by Levenberg-Marquardt for cotton crop classification. To experiment the proposed method, five LISS-III satellite images was taken and the experimentation was done with six vegetation indices such as Simple Ratio, Normalized Difference Vegetation Index, Enhanced Vegetation Index, Green Atmospherically Resistant Vegetation Index, Wide-Dynamic Range Vegetation Index, Green Chlorophyll Index. Along with these indices, Green Leaf Area Index is also considered for investigation. From the research outcome, Green Atmospherically Resistant Vegetation Index outperformed with all other indices by reaching the average accuracy value of 95.21%.Keywords: Fuzzy C-Means clustering (FCM), neural network, Levenberg-Marquardt (LM) algorithm, vegetation indices
Procedia PDF Downloads 310914 Synthesis, Crystallography and Anti-TB Activity of Substituted Benzothiazole Analogues
Authors: Katharigatta N. Venugopala, Melendhran Pillay, Bander E. Al-Dhubiab
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Tuberculosis (TB) infection is caused mainly by Mycobacterium tuberculosis (MTB) and it is one of the most threatening and wide spread infectious diseases in the world. Benzothiazole derivatives are found to have diverse chemical reactivity and broad spectrum of pharmacological activity. Some of the important pharmacological activities shown by the benzothiazole analogues are antitumor, anti-inflammatory, antimicrobial, anti-tubercular, anti-leishmanial, anticonvulsant and anti-HIV properties. Keeping all these facts in mind in the present investigation it was envisaged to synthesize a series of novel {2-(benzo[d]-thiazol-2-yl-methoxy)-substitutedaryl}-(substitutedaryl)-methanones (4a-f) and characterize by IR, NMR (1H and 13C), HRMS and single crystal x-ray studies. The title compounds are investigated for in vitro anti-tubercular activity against two TB strains such as H37Rv (ATCC 25177) and MDR-MTB (multi drug resistant MTB resistant to Isoniazid, Rifampicin and Ethambutol) by agar diffusion method. Among the synthesized compounds in the series, test compound {2-(benzo[d]thiazol-2-yl-methoxy)-5-fluorophenyl}-(4-chlorophenyl)-methanone (2c) was found to exhibit significant activity with MICs of 1 µg/mL and 2 µg/mL against H37Rv and MDR-MTB, respectively when compared to standard drugs. Single crystal x-ray studies was used to study intra and intermolecular interactions, including polymorphism behavior of the test compounds, but none of the compounds exhibited polymorphism behavior.Keywords: benzothiazole analogues, characterization, crystallography, anti-TB activity
Procedia PDF Downloads 272913 Antibiotic Resistance of Enterococci Isolated from Raw Cow Milk
Authors: Margita Čanigová, Jana Račková, Miroslav Kročko, Viera Ducková, Vladimíra Kňazovická
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The aim of the study was to test the milk samples in terms of enterococci presence and their counts. Tested samples were as follows: raw cow milk, raw cow milk stored at 10°C for 16 hours and milk pasteurised at 72°C for 15 seconds. The typical colonies were isolated randomly and identified by classical biochemical test - EN-COCCUS test (Lachema, CR) and by PCR. Isolated strains were tested in terms of antibiotic resistance by well diffusion method. Examined antibiotics were: vancomycin (30 μg/disc), gentamicin (120 μg/disc), erythromycin (15 μg/disc), teicoplanine (30 μg/disc), ampicillin (10 μg/disc) and tetracycline (30 μg/disc). Average value of enterococci counts in raw milk cistern samples (n=30) was 8.25 ± 1.37 ×103 CFU/cm3. Storage tank milk samples (n=30) showed an increase (P > 0.05) and average value was 9.16 ± 1.49 × 103 CFU/cm3. Occurrence of enterococci in pasteurized milk (n=30) was sporadic and their counts were mostly below 10 CFU/cm3. Overall, 96 enterococci strains were isolated. In samples of raw cow milk and stored raw cow milk, Enterococcus faecalis was a dominant species (58.1% and 71.7%, respectively), followed by E. faecium (16.3% and 0%, respectively). Enterococcus mundtii, E. casseliflavus, E. durans and E. gallinarum were isolated, too. Resistances to ampicillin, erythromycin, gentamicin, tetracycline and vancomycin were found in 7.29%, 3.13%, 4.00%, 13.54% and 10.42% of isolated enterococci strains, respectively. Resistance to teicoplanine was not found in any isolated strain. All Vancomycin-Resistant Enterococci (VRE) belonged to E. faecalis. Obtained results confirmed that raw milk is a potential risk of enterococci resistant to antibiotics transmission into the food chain.Keywords: antibiotic resistance, enterococci, milk, biosystems engineering
Procedia PDF Downloads 372912 A Comparative Study: Comparison of Two Different Fluorescent Stains -Auramine and Rhodamine- with Ehrlich-Ziehl-Neelsen, Kinyoun Staining, and Culture in the Determination of Acid Resistant Bacilli
Authors: Recep Keşli, Hayriye Tokay, Cengiz Demir, İsmail Ceyhan
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Objective: In many countries, tuberculosis (TB) is still one of the most important diseases. Tuberculosis is among top 10 causes of death worldwide. The early diagnosis of active tuberculosis still depends on the presence of acid resistant bacilli (ARB) in stained smears. In this study, we aimed to investigate the diagnostic performances of Erlich Ziehl Neelsen (EZN), Kinyoun and two different fluorescent stains. Methods: The specimens were obtained from the patients who applied to Chest Diseases Departments of Ankara Atatürk Chest Diseases and Thoracic Surgery Training and Research Hospital, and Afyon Kocatepe University, ANS Research and Practice Hospital. The study was carried out in the Medical Microbiology Laboratory, School of Medicine, Afyon Kocatepe University. All the non-sterile specimens were homogenized and decontaminated according to the EUCAST instructions. Samples were inoculated onto the Löwenstein-Jensen agars (bio-Merieux Marcy l'Etoile, France) and then incubated at 37˚C, for 40 days. Four smears were prepared from each specimen. Slides were stained with commercial EZN (BD, Sparks, USA), Kinyoun (SALUBRIS Istanbul, Turkey), Auramine (SALUBRIS Istanbul, Turkey) and Rhodamine (SALUBRIS Istanbul, Turkey) kit. While EZN and Kinyoun stainings were examined by light microscope, Auramine and Rhodamine slides were examined by fluorescence microscopy. Results: A total of 158 respiratory system samples (sputum, broncho alveolar lavage fluid…etc) were enrolled into the study. A hundred and two of the samples that processed were found as culture positive. The sensitivity, specificity, positive predictive, and negative predictive values were detected as 100%, 67.5%, 73.5%, and 100% for EZN, 100%, 70.9%, 77.4%, and 100% for Kinyoun, 100%,77.8%, 84.3%, 100% for Auramine, and 100%, 80% , 86.3%, and 100% for Rhodamine respectively. Conclusions: According to our study auramine and rhodamine staining methods showed the best diagnostic performance among the four investigated staining methods. In conclusion, the fluorochrome staining method may be accepted as the most reliable, rapid and useful method for diagnosis of the mycobacterial infections truly.Keywords: acid resistant bacilli (ARB), auramine, Ehrlich-Ziehl-Neelsen (EZN), Kinyoun, Rhodamine
Procedia PDF Downloads 272911 Synthesis, Antibacterial Activities, and Synergistic Effects of Novel Juglone and Naphthazarin Derivatives Against Clinical Methicillin-Resistant Staphylococcus aureus Strains
Authors: Zohra Benfodda, Valentin Duvauchelle, Chaimae Majdi, David Bénimélis, Catherine Dunyach-Remy, Patrick Meffre
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New antibiotics are necessary to treat microbial pathogens, especially ESKAPE pathogens that are becoming increasingly resistant to available treatment. Despite the medical need, the number of newly approved drugs continues to decline. The majority of antibiotics under clinical development are natural products or derivatives thereof. 43 juglone/naphthazarin derivatives were synthesized using Minisci-type direct C–H alkylation and evaluated for their antibacterial properties against various clinical and reference Gram-positive MSSA, clinical Gram-positive MRSA. Different compounds of the synthesized series showed promising activity against clinical and reference MSSA (MIC: 1–8 μg/ml) and good efficacy against clinical MRSA (MIC: 2–8 μg/ml) strains. The synergistic effects of active compounds were evaluated with reference antibiotics (vancomycin and cloxacillin), and it was found that the antibiotic combination with those active compounds efficiently enhanced the antimicrobial activity and consequently the MIC values of reference antibiotics were lowered up to 1/16th of the original MIC. These synthesized compounds did not present hemolytic activity on sheep red blood cells. In addition to the in silico prediction of ADME profile parameter which is promising and encouraging for further development.Keywords: juglone, naphthazarin, antibacterial, clinical MRSA, synergistic studies, MIC determination
Procedia PDF Downloads 117910 Screening the Best Integrated Pest Management Treatments against Helicoverpa armigera
Authors: Ajmal Khan Kassi, Humayun Javed, Tariq Mukhtar
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The research was conducted to screen out resistance and susceptibility of okra varieties against Helicoverpa armigera under field conditions 2016. In this experiment, the different management practices viz. release Trichogramma chilonis, hoeing, and weeding, clipping, and lufenuron were tested individually and with all possible combinations for the controlling of American bollworm at 3 diverse localities viz. University research farm Koont, National Agriculture Research Centre (NARC) and farmer field Taxila by using resistant variety Arka Anamika. All the treatment combinations regarding damage of shoot and fruit showed significant results. The minimum fruit infestation, i.e., 3.20% and 3.58% was recorded with combined treatment (i.e., T. chilonis + hoeing + weeding + lufenuron) in two different localities. The minimum shoot infestation, i.e., 7.18%, 7.08%, and 6.85% was also observed with (T. chilonis + hoeing + weeding + lufenuron) combined treatment at all three different localities. The above-combined treatment (T. chilonis + hoeing + weeding + lufenuron) also resulted in maximum yield at NARC and Taxila, i.e., 57.67 and 62.66 q/ha respectively. On the basis of combined treatment (i.e., T. chilonis + hoeing + weeding + lufenuron) in three different localities, Arka Anamika variety proved to be comparatively resistant against H. armigera. So this variety is recommended for the cultivation in Pothwar region to get maximum yield and minimum losses against H. armigera.Keywords: okra, screening, combine treatment, Helicoverpa armigera
Procedia PDF Downloads 151909 Exploring Penicillin Resistance in Gonococcal Penicillin Binding Protein-2: Molecular Docking and Ligand Interaction Analysis
Authors: Sinethemba Yakobi, Lindiwe Zuma, Ofentse Pooe
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Gonococcal infections present a notable public health issue, and the major approach for treatment involves using β-lactam antibiotics that specifically target penicillin-binding protein 2 (PBP2) in Neisseria gonorrhoeae. This study examines the influence of flavonoids, namely rutin, on the structural changes of PBP2 in both penicillin-resistant (FA6140) and penicillin-susceptible (FA19) strains. The research clarifies the structural effects of particular mutations, such as inserting an aspartate residue at position 345 (Asp-345a) in the PBP2 protein. The strain FA6140, which is resistant to penicillin, shows specific changes that lead to a decrease in penicillin binding. These mutations, namely P551S and F504L, significantly impact the pace at which acylation occurs and the stability of the strain under high temperatures. Molecular docking analyses investigate the antibacterial activities of rutin and other phytocompounds, emphasizing its exceptional binding affinity and potential as an inhibitor of PBP2. Quercetin and protocatechuic acid have encouraging antibacterial effectiveness, with quercetin displaying characteristics similar to those of drugs. Molecular dynamics simulations offer a detailed comprehension of the interactions between flavonoids and PBP2, highlighting rutin's exceptional antioxidant effects and strong affinity for the substrate binding site. The study's wider ramifications pertain to the pressing requirement for antiviral treatments in the context of the ongoing COVID-19 epidemic. Flavonoids have a strong affinity for binding to PBP2, indicating their potential as inhibitors to impair cell wall formation in N. gonorrhoeae. Ultimately, this study provides extensive knowledge on the interactions between proteins and ligands, the dynamics of the structure, and the ability of flavonoids to combat penicillin-resistant N. gonorrhoeae bacteria. The verified simulation outcomes establish a basis for creating potent inhibitors and medicinal therapies to combat infectious illnesses.Keywords: phytochemicals, penicillin-binding protein 2, gonococcal infection, ligand-protein interaction, binding energy, neisseria gonorrhoeae FA19, neisseria gonorrhoeae FA6140, flavonoids
Procedia PDF Downloads 61908 Effect of Different Nitrogen Level on Vegetative Growth of Maize Variety (Zea Mays)
Authors: Tegene Nigussie
Abstract:
Introduction: Maize is the most domesticated of all the field crops. Wild maize has not been found to date and there has been much speculation on its origin. Regardless of the validity of different theories, it is generally agreed that the center of origin of maize is Central America, primarily Mexico and the Caribbean. Maize in Africa is of a recent introduction although data suggest that it was present in Nigeria even before Columbus voyages. After being taken to Europe in 1493, maize was introduced to Africa and distributed through the continent by different routes. Maize is an important cereal crop in Ethiopia. In general, it is the primarily stable food, and rural households show a strong preference. For human food, the important constituents of grain are carbohydrates (starch and sugars), protein, fat or oil (in the embryo) and minerals. About 75 percent of the kernel is starch, a range of 60.80 percent, but low protein content (8-15). In Ethiopia, the introduction of modern farming techniques appears to be a priority. However, the adoption of modern inputs by peasant farmers is found to be very slow; for example, the adoption rate of fertilizer, an input that is relatively adopted, is very slow. The difference socio economic factors lied behind the low rate of technology adoption, including price &marketing input. Objective: The objective of this study is to determine the optimum application rate or level of different nitrogen fertilizers for the vegetative growth of maize and to identify the effect of different nitrogen rates on the growth and development of maize. Methods: The vegetative parameter (above ground) measurement from five plants randomly sampled from the middle rows of each plot. Results: The interaction of nitrogen and maize variety showed a significant at (p<0.01) effect on plant height, with the application of 60kg/ha and BH140 maize variety in combination and root length with the application of 60kg/ha of nitrogen and BH140 variety of maize. The highest mean (12.33) of the number of leaves per plant and mean (7.1) of the number of nodes per plant can be used as an alternative for better vegetative growth of maize. Conclusion: Maize is one of the most popular and cultivated crops in Ethiopia. The study was conducted to investigate the best dosage of nitrogen for vegetative growth, yield, and better quality of maize variety and to recommend a level of nitrogen rate and the best variety adaptable to the specific soil condition or area.Keywords: parameter, chlorosis, germination, flood, sesbania, cultivar
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