Search results for: bacterial inoculum
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1156

Search results for: bacterial inoculum

796 Isolation and Screening of Antagonistic Bacteria against Wheat Pathogenic Fungus Tilletia indica

Authors: Sugandha Asthana, Geetika Vajpayee, Pratibha Kumari, Shanthy Sundaram

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An economically important disease of wheat in North Western region of India is Karnal Bunt caused by smut fungus Tilletia indica. This fungal pathogen spreads by air, soil and seed borne sporodia at the time of flowering, which ultimately leads to partial bunting of wheat kernels with fishy odor and taste to wheat flour. It has very serious effects due to quarantine measures which have to be applied for grain exports. Chemical fungicides such as mercurial compounds and Propiconazole applied to the control of Karnal bunt have been only partially successful. Considering the harmful effects of chemical fungicides on man as well as environment, many countries are developing biological control as the superior substitute to chemical control. Repeated use of fungicides can be responsible for the development of resistance in fungal pathogens against certain chemical compounds. The present investigation is based on the isolation and evaluation of antifungal properties of some isolated (from natural manure) and commercial bacterial strains against Tilletia indica. Total 23 bacterial isolates were obtained and antagonistic activity of all isolates and commercial bacterial strains (Bacillus subtilis MTCC8601, Bacillus pumilus MTCC 8743, Pseudomonas aeruginosa) were tested against T. indica by dual culture plate assay (pour plate and streak plate). Test for the production of antifungal volatile organic compounds (VOCs) by antagonistic bacteria was done by sealed plate method. Amongst all s1, s3, s5, and B. subtilis showed more than 80% inhibition. Production of extracellular hydrolytic enzymes such as protease, beta 1, 4 glucanase, HCN and ammonia was studied for confirmation of antifungal activity. s1, s3, s5 and B. subtilis were found to be the best for protease activity and s5 and B. subtilis for beta 1, 4 glucanase activity. Bacillus subtilis was significantly effective for HCN whereas s3, s5 and Bacillus subtilis for ammonia production. Isolates were identified as Pseudomonas aeruginosa (s1) and B. licheniformis (s3, s5) by various biochemical assays and confirmed by16s rRNA sequencing. Use of microorganisms or their secretions as biocontrol agents to avoid plant diseases is ecologically safe and may offer long term of protection to crop. The above study reports the promising effects of these strains in better pathogen free crop production and quality maintenance as well as prevention of the excessive use of synthetic fungicides.

Keywords: antagonistic, antifungal, biocontrol, Karnal bunt

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795 Antimicrobial Activity of Some Plant Extracts against Clinical Pathogen and Candida Species

Authors: Marwan Khalil Qader, Arshad Mohammad Abdullah

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Antimicrobial resistance is a major cause of significant morbidity and mortality globally. Seven plant extracts (Plantago mediastepposa, Quercusc infectoria, Punic granatum, Thymus lcotschyana, Ginger officeinals, Rhus angustifolia and Cinnamon) were collected from different regions of Kurdistan region of Iraq. These plants’ extracts were dissolved in absolute ethanol and distillate water, after which they were assayed in vitro as an antimicrobial activity against Candida tropicalis, Candida albicanus, Candida dublinensis, Candida krusei and Candida glabrata also against 2 Gram-positive (Bacillus subtilis and Staphylococcus aureus) and 3 Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Klebsilla pneumonia). The antimicrobial activity was determined in ethanol extracts and distilled water extracts of these plants. The ethanolic extracts of Q. infectoria showed the maximum activity against all species of Candida fungus. The minimum inhibition zone of the Punic granatum ethanol extracts was 0.2 mg/ml for all microorganisms tested. Klebsilla pneumonia was the most sensitive bacterial strain to Quercusc infectoria and Rhus angustifolia ethanol extracts. Among both Gram-positive and Gram-negative bacteria tested with MIC of 0.2 mg/ml, the minimum inhibition zone of Ginger officeinals D. W. extracts was 0.2 mg/mL against Pseudomonas aeruginosa and Klebsilla pneumonia. The most sensitive bacterial strain to Thymus lcotschyana and Plantago mediastepposa D.W. extracts was S. aureus and E. coli.

Keywords: antimicrobial activity, pathogenic bacteria, plant extracts, chemical systems engineering

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794 Sulfur-Containing Diet Shift Hydrogen Metabolism and Reduce Methane Emission and Modulated Gut Microbiome in Goats

Authors: Tsegay Teklebrhan Gebremariam, Zhiliang, Arjan Jonker

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The study investigated that using corn gluten (CG) instead of cornmeal (CM) increased dietary sulfur shifted H₂ metabolism from methanogenesis to alternative sink and modulated microbiome in the rumen as well as hindgut segments of goats. Ruminal fermentation, CH₄ emissions and microbial abundance in goats (n = 24). The experiment was performed using a randomized block design with two dietary treatments (CM and CG with 400 g/kg DM each). Goats in CG increased sulfur, NDF and CP intake and decreased starch intake as compared with those in CM. Goats that received CG diet had decreased dissolved hydrogen (dH₂) (P = 0.01) and dissolved methane yield and emission (dCH₄) (P = 0.001), while increased dH₂S both in the rumen and hindgut segments than those fed CM. Goats fed CG had higher (p < 0.01) gene copies of microbiota and cellulolytic bacteria, whereas starch utilizing bacterial species were less in the rumen and hindgut than those fed CM. Higher (P < 0.05) methanogenic diversity and abundances of Methanimicrococcus and Methanomicrobium were observed in goats that consumed CG, whilst containing lower Methanobrevibacter populations than those receiving CM. The study suggested that goats fed corn gluten improved the gene copies of microbiota and fibrolytic bacterial species while reducing starch utilizing species in the rumen and hindgut segments as compared with that fed cornmeal. Goats consuming corn gluten had a more enriched methanogenic diversity and reduced Methanobrevibacter, a contributor to CH₄ emissions, as compared with goats fed CM. Corn gluten could be used as an alternative feed to decrease the enteric CH₄ emission in ruminant production.

Keywords: dissolved gasses, methanogenesis, microbial community, metagenomics

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793 Effect of Ethanol and Betadine on the Preformed Biofilm of Staphylococcus Aureus Isolated from Urinary Catheter

Authors: Kara Terki Ibtissem, Hassaine Hafida, Bellifa Samia

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Introduction: Staphylococcus aureus is one of the species that are most frequently isolated from urinary catheters. The ability to produce a biofilm is an important step in the pathogenesis of these staphylococci; biofilm formation is strongly dependent on the environmental conditions it also depends on the different parameters these biofilms are subjected to. Antiseptics, including ethanol and betadine, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that disinfectants may enhance biofilm production in Staphylococci. Methods: In this study, 48 staphylococcus aureus isolated from urinary catheters at the University Hospital Center of Sidi Bel Abbes (in Northwestern Algeria) were analyzed to detect the formation of biofilm by culture on Red Congo Agar (RCA), the Tube Method (TM) and tissue Culture Plate (TCP) techniques, this last was also used to investigate the effect of ethanol and Betadine on the preformed biofilm In a second time to know which environment is most favorable to the formation of the biofilm we perform a statistical test based on the student test by the software R. Results: It has been found that 23 strains produced a bacterial slime on the Congo red medium, 5 strains produced a biofilm by the tube method, 2 of which are highly productive. In addition, 7 strains produced a biofilm on polystyrene micro-plates; this number was higher in the presence of ethanol 70% and ethanol 90% with 19 and 11 biofilm-producing strains, respectively. On the other hand, no biofilm was formed in the presence of Betadine. Conclusion: It is important to examine the response of biofilms following an imposed external constraint, such as disinfectants, in order to develop new strategies to combat bacterial biofilms but also to better control their formation.

Keywords: staphylococcus aureus, biofilm, urinary catheter, ethanol

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792 Varietal Screening of Watermelon against Powdery Mildew Disease and Its Management

Authors: Asim Abbasi, Amer Habib, Sajid Hussain, Muhammad Sufyan, Iqra, Hasnain Sajjad

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Except for few scattered cases, powdery mildew disease was not a big problem for watermelon in the past but with the outbreaks of its pathotypes, races 1W and 2W, this disease becomes a serious issue all around the globe. The severe outbreak of this disease also increased the rate of fungicide application for its proper management. Twelve varieties of watermelon were screened in Research Area of Department of Plant pathology, University of Agriculture, Faisalabad to check the incidence of powdery mildew disease. Disease inoculum was prepared and applied with the help of foliar spray method. Fungicides and plants extracts were also applied after the disease incidence. Percentage leaf surface area diseased was assessed visually with a modified Horsfall-Barratt scale. The results of the experiment revealed that among all varieties, WT2257 and Zcugma F1 were highly resistant showing less than 5% disease incidence while Anar Kali and Sugar baby were highly susceptible with disease incidence of more than 65%. Among botanicals neem extract gave best results with disease incidence of less than 20%. Besides neem, all other botanicals also gave significant control of powdery mildew disease than the untreated check. In case of fungicides, Gemstar showed least disease incidence i.e. < 10%, however besides control maximum disease incidence was observed in Curzate (> 30%).

Keywords: botanicals, fungicides, pathotypes, powdery mildew

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791 Bacterial Exposure and Microbial Activity in Dental Clinics during Cleaning Procedures

Authors: Atin Adhikari, Sushma Kurella, Pratik Banerjee, Nabanita Mukherjee, Yamini M. Chandana Gollapudi, Bushra Shah

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Different sharp instruments, drilling machines, and high speed rotary instruments are routinely used in dental clinics during dental cleaning. Therefore, these cleaning procedures release a lot of oral microorganisms including bacteria in clinic air and may cause significant occupational bioaerosol exposure risks for dentists, dental hygienists, patients, and dental clinic employees. Two major goals of this study were to quantify volumetric airborne concentrations of bacteria and to assess overall microbial activity in this type of occupational environment. The study was conducted in several dental clinics of southern Georgia and 15 dental cleaning procedures were targeted for sampling of airborne bacteria and testing of overall microbial activity in settled dusts over clinic floors. For air sampling, a Biostage viable cascade impactor was utilized, which comprises an inlet cone, precision-drilled 400-hole impactor stage, and a base that holds an agar plate (Tryptic soy agar). A high-flow Quick-Take-30 pump connected to this impactor pulls microorganisms in air at 28.3 L/min flow rate through the holes (jets) where they are collected on the agar surface for approx. five minutes. After sampling, agar plates containing the samples were placed in an ice chest with blue ice and plates were incubated at 30±2°C for 24 to 72 h. Colonies were counted and converted to airborne concentrations (CFU/m3) followed by positive hole corrections. Most abundant bacterial colonies (selected by visual screening) were identified by PCR amplicon sequencing of 16S rRNA genes. For understanding overall microbial activity in clinic floors and estimating a general cleanliness of the clinic surfaces during or after dental cleaning procedures, ATP levels were determined in swabbed dust samples collected from 10 cm2 floor surfaces. Concentration of ATP may indicate both the cell viability and the metabolic status of settled microorganisms in this situation. An ATP measuring kit was used, which utilized standard luciferin-luciferase fluorescence reaction and a luminometer, which quantified ATP levels as relative light units (RLU). Three air and dust samples were collected during each cleaning procedure (at the beginning, during cleaning, and immediately after the procedure was completed (n = 45). Concentrations at the beginning, during, and after dental cleaning procedures were 671±525, 917±1203, and 899±823 CFU/m3, respectively for airborne bacteria and 91±101, 243±129, and 139±77 RLU/sample, respectively for ATP levels. The concentrations of bacteria were significantly higher than typical indoor residential environments. Although an increasing trend for airborne bacteria was observed during cleaning, the data collected at three different time points were not significantly different (ANOVA: p = 0.38) probably due to high standard deviations of data. The ATP levels, however, demonstrated a significant difference (ANOVA: p <0.05) in this scenario indicating significant change in microbial activity on floor surfaces during dental cleaning. The most common bacterial genera identified were: Neisseria sp., Streptococcus sp., Chryseobacterium sp., Paenisporosarcina sp., and Vibrio sp. in terms of frequencies of occurrences, respectively. The study concluded that bacterial exposure in dental clinics could be a notable occupational biohazard, and appropriate respiratory protections for the employees are urgently needed.

Keywords: bioaerosols, hospital hygiene, indoor air quality, occupational biohazards

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790 Thrombocytopenia and Prolonged Prothrombin Time in Neonatal Septicemia

Authors: Shittu Bashirat, Shittu Mujeeb, Oluremi Adeolu, Orisadare Olayiwola, Jikeme Osameke, Bello Lateef

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Septicemia in neonates refers to generalized bacterial infection documented by positive blood culture in the first 28 days of life and is one of the leading causes of neonatal mortality in sub-Sahara Africa. Thrombocytopenia in newborns is a result of increased platelet consumption; sepsis was found to be the most common risk factor. The objective of the study was to determine if there are organism-specific platelet responses among the 2 groups of bacterial agents: Gram-positive and Gram-negative bacteria, and also to examine the association of platelet count and prothrombin time with neonatal septicemia. 232 blood samples were collected for this study. The blood culture was performed using Bactec 9050, an instrumented blood culture system. The platelet count and prothrombin time were performed using Abacus Junior 5 hematology analyzer and i-STAT 1 analyzer respectively. Of the 231 neonates hospitalized with clinical sepsis, blood culture reports were positive in 51 cases (21.4%). Klebsiella spp. (35.3%) and Staphylococcus aureus (27.5%) were the most common Gram-negative and Gram-positive isolates respectively. Thrombocytopenia was observed in 30 (58.8%) of the neonates with septicemia. Of the 9 (17.6%) patients with severe thrombocytopenia, seven (77.8%) had Klebsiella spp. septicemia. Out of the 21(63.6%) of thrombocytopenia produced by Gram-negative isolate, 17 (80.9) had increased prothrombin time. In conclusion, Gram-negative organisms showed the highest cases of severe thrombocytopenia and prolonged PT. This study has helped to establish a disturbance in hemostatic systems in neonates with septicemia. Further studies, however, may be required to assess other hemostasis parameters in order to understand their interaction with the infectious organisms in neonates.

Keywords: neonates, septicemia, thrombocytopenia, prolonged prothrombin time, platelet count

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789 Metabolic and Phylogenetic Profiling of Rhizobium leguminosarum Strains Isolated from NZ Soils of Varying pH

Authors: Anish Shah, Steve A. Wakelin, Derrick Moot, Aurélie Laugraud, Hayley J. Ridgway

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A mixed pasture system of ryegrass-clover is used in New Zealand, where clovers are generally inoculated with commercially available strains of rhizobia. The community of rhizobia living in the soil and the way in which they interact with the plant are affected by different biotic and abiotic factors. In general, bacterial richness and diversity in soil varies by soil pH. pH also affects cell physiology and acts as a master variable that controls the wider soil physiochemical conditions such as P availability, Al release and micronutrient availability. As such, pH can have both primary and secondary effects on soil biology and processes. The aim of this work was to investigate the effect of soil pH on the genetic diversity and metabolic profile of Rhizobium leguminosarum strains nodulating clover. Soils were collected from 12 farms across New Zealand which had a pH(water) range of between 4.9 and 7.5, with four acidic (pH 4.9 – 5.5), four ‘neutral’ (5.8 – 6.1) and four alkaline (6.5 – 7.5) soils. Bacteria were recovered from nodules of Trifolium repens (white clover) and T. subterraneum (subterranean clover) grown in the soils. The strains were cultured and screened against a range of pH-amended media to demonstrate whether they were adapted to pH levels similar to their native soils. The strains which showed high relative growth at a given pH (~20% of those isolated) were selected for metabolic and taxonomic profiling. The Omnilog (Biolog Inc., Hayward, CA) phenotype array was used to perform assays on carbon (C) utilisation for selected strains. DNA was extracted from the strains which had differing C utilisation profiles and PCR products for both forward and reverse primers were sequenced for the following genes: 16S rRNA, recA, nodC, nodD and nifH (symbiotic).

Keywords: bacterial diversity, clover, metabolic and taxonomic profiling, pH adaptation, rhizobia

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788 Blood Microbiome in Different Metabolic Types of Obesity

Authors: Irina M. Kolesnikova, Andrey M. Gaponov, Sergey A. Roumiantsev, Tatiana V. Grigoryeva, Dilyara R. Khusnutdinova, Dilyara R. Kamaldinova, Alexander V. Shestopalov

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Background. Obese patients have unequal risks of metabolic disorders. It is accepted to distinguish between metabolically healthy obesity (MHO) and metabolically unhealthy obesity (MUHO). MUHO patients have a high risk of metabolic disorders, insulin resistance, and diabetes mellitus. Among the other things, the gut microbiota also contributes to the development of metabolic disorders in obesity. Obesity is accompanied by significant changes in the gut microbial community. In turn, bacterial translocation from the intestine is the basis for the blood microbiome formation. The aim was to study the features of the blood microbiome in patients with various metabolic types of obesity. Patients, materials, methods. The study included 116 healthy donors and 101 obese patients. Depending on the metabolic type of obesity, the obese patients were divided into subgroups with MHO (n=36) and MUHO (n=53). Quantitative and qualitative assessment of the blood microbiome was based on metagenomic analysis. Blood samples were used to isolate DNA and perform sequencing of the variable v3-v4 region of the 16S rRNA gene. Alpha diversity indices (Simpson index, Shannon index, Chao1 index, phylogenetic diversity, the number of observed operational taxonomic units) were calculated. Moreover, we compared taxa (phyla, classes, orders, and families) in terms of isolation frequency and the taxon share in the total bacterial DNA pool between different patient groups. Results. In patients with MHO, the characteristics of the alpha-diversity of the blood microbiome were like those of healthy donors. However, MUHO was associated with an increase in all diversity indices. The main phyla of the blood microbiome were Bacteroidetes, Firmicutes, Proteobacteria, and Actinobacteria. Cyanobacteria, TM7, Thermi, Verrucomicrobia, Chloroflexi, Acidobacteria, Planctomycetes, Gemmatimonadetes, and Tenericutes were found to be less significant phyla of the blood microbiome. Phyla Acidobacteria, TM7, and Verrucomicrobia were more often isolated in blood samples of patients with MUHO compared with healthy donors. Obese patients had a decrease in some taxonomic ranks (Bacilli, Caulobacteraceae, Barnesiellaceae, Rikenellaceae, Williamsiaceae). These changes appear to be related to the increased diversity of the blood microbiome observed in obesity. An increase of Lachnospiraceae, Succinivibrionaceae, Prevotellaceae, and S24-7 was noted for MUHO patients, which, apparently, is explained by a magnification in intestinal permeability. Conclusion. Blood microbiome differs in obese patients and healthy donors at class, order, and family levels. Moreover, the nature of the changes is determined by the metabolic type of obesity. MUHO linked to increased diversity of the blood microbiome. This appears to be due to increased microbial translocation from the intestine and non-intestinal sources.

Keywords: blood microbiome, blood bacterial DNA, obesity, metabolically healthy obesity, metabolically unhealthy obesity

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787 A Study on the Microbilogical Profile and Antibiotic Sensitivity Pattern of Bacterial Isolates Causing Urinary Tract Infection in Intensive Care Unit Patients in a Tertiary Care Hospital in Eastern India

Authors: Pampita Chakraborty, Sukumar Mukherjee

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The study was done to determine the microbiological profile and changing pattern of the pathogens causing UTI in the ICU patients. All the patients admitted to the ICU with urinary catheter insertion for more than 48hours were included in the study. Urine samples were collected in a sterile container with aseptic precaution using disposable syringe and was processed as per standards. Antimicrobial susceptibility test was done by Disc Diffusion method as per CLSI guidelines. A total of 100 urine samples were collected from ICU patients, out of which 30% showed significant bacterial growth and 7% showed growth of candida spp. Prevalence of UTI was more in female (73%) than male (27.%). Gram-negative bacilli 26(86.67%) were more common in our study followed by gram-positive cocci 4(13.33%). The most common uropathogens isolated were Escherichia coli 14 (46.67%), followed by Klebsiella spp 7(23.33%), Staphylococcus aureus 4(13.33%), Acinetobacter spp 3(10%), Enterococcus faecalis 1(3.33%) and Pseudomonas aeruginosa 1(3.33%). Most of the Gram-negative bacilli were sensitive to amikacin (80%) and nitrofurantoin (80%), where as all gram-positive organisms were sensitive to Vancomycin. A large number ESBL producers were also observed in this study. The study finding showed that E.coli is the predominant pathogen and has increasing resistance pattern to the commonly used antibiotics. The study proposes that the adherence to antibiotic policy is the key ingredients for successful outcome in ICU patients and also emphasizes that repeated evaluation of microbial characteristics and continuous surveillance of resistant bacteria is required for selection of appropriate antibiotic therapy.

Keywords: antimicrobial sensitivity, intensive care unit, nosocomial infection, urinary tract infection

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786 Bio Energy from Metabolic Activity of Bacteria in Plant and Soil Using Novel Microbial Fuel Cells

Authors: B. Samuel Raj, Solomon R. D. Jebakumar

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Microbial fuel cells (MFCs) are an emerging and promising method for achieving sustainable energy since they can remove contaminated organic matter and simultaneously generate electricity. Our approach was driven in three different ways like Bacterial fuel cell, Soil Microbial fuel cell (Soil MFC) and Plant Microbial fuel cell (Plant MFC). Bacterial MFC: Sulphate reducing bacteria (SRB) were isolated and identified as the efficient electricigens which is able to produce ±2.5V (689mW/m2) and it has sustainable activity for 120 days. Experimental data with different MFC revealed that high electricity production harvested continuously for 90 days 1.45V (381mW/m2), 1.98V (456mW/m2) respectively. Biofilm formation was confirmed on the surface of the anode by high content screening (HCS) and scanning electron Microscopic analysis (SEM). Soil MFC: Soil MFC was constructed with low cost and standard Mudwatt soil MFC was purchased from keegotech (USA). Vermicompost soil (V1) produce high energy (± 3.5V for ± 400 days) compared to Agricultural soil (A1) (± 2V for ± 150 days). Biofilm formation was confirmed by HCS and SEM analysis. This finding provides a method for extracting energy from organic matter, but also suggests a strategy for promoting the bioremediation of organic contaminants in subsurface environments. Our Soil MFC were able to run successfully a 3.5V fan and three LED continuously for 150 days. Plant MFC: Amaranthus candatus (P1) and Triticum aestivium (P2) were used in Plant MFC to confirm the electricity production from plant associated microbes, four uniform size of Plant MFC were constructed and checked for energy production. P2 produce high energy (± 3.2V for 40 days) with harvesting interval of two times and P1 produces moderate energy without harvesting interval (±1.5V for 24 days). P2 is able run 3.5V fan continuously for 10days whereas P1 needs optimization of growth conditions to produce high energy.

Keywords: microbial fuel cell, biofilm, soil microbial fuel cell, plant microbial fuel cell

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785 Effect of Phosphorus Solubilizing Bacteria on Yield and Seed Quality of Camelina (Camelina sativa L.) under Drought Stress

Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

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New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

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784 Enhancing Algal Bacterial Photobioreactor Efficiency: Nutrient Removal and Cost Analysis Comparison for Light Source Optimization

Authors: Shahrukh Ahmad, Purnendu Bose

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Algal-Bacterial photobioreactors (ABPBRs) have emerged as a promising technology for sustainable biomass production and wastewater treatment. Nutrient removal is seldom done in sewage treatment plants and large volumes of wastewater which still have nutrients are being discharged and that can lead to eutrophication. That is why ABPBR plays a vital role in wastewater treatment. However, improving the efficiency of ABPBR remains a significant challenge. This study aims to enhance ABPBR efficiency by focusing on two key aspects: nutrient removal and cost-effective optimization of the light source. By integrating nutrient removal and cost analysis for light source optimization, this study proposes practical strategies for improving ABPBR efficiency. To reduce organic carbon and convert ammonia to nitrates, domestic wastewater from a 130 MLD sewage treatment plant (STP) was aerated with a hydraulic retention time (HRT) of 2 days. The treated supernatant had an approximate nitrate and phosphate values of 16 ppm as N and 6 ppm as P, respectively. This supernatant was then fed into the ABPBR, and the removal of nutrients (nitrate as N and phosphate as P) was observed using different colored LED bulbs, namely white, blue, red, yellow, and green. The ABPBR operated with a 9-hour light and 3-hour dark cycle, using only one color of bulbs per cycle. The study found that the white LED bulb, with a photosynthetic photon flux density (PPFD) value of 82.61 µmol.m-2 .sec-1 , exhibited the highest removal efficiency. It achieved a removal rate of 91.56% for nitrate and 86.44% for phosphate, surpassing the other colored bulbs. Conversely, the green LED bulbs showed the lowest removal efficiencies, with 58.08% for nitrate and 47.48% for phosphate at an HRT of 5 days. The quantum PAR (Photosynthetic Active Radiation) meter measured the photosynthetic photon flux density for each colored bulb setting inside the photo chamber, confirming that white LED bulbs operated at a wider wavelength band than the others. Furthermore, a cost comparison was conducted for each colored bulb setting. The study revealed that the white LED bulb had the lowest average cost (Indian Rupee)/light intensity (µmol.m-2 .sec-1 ) value at 19.40, while the green LED bulbs had the highest average cost (INR)/light intensity (µmol.m-2 .sec-1 ) value at 115.11. Based on these comparative tests, it was concluded that the white LED bulbs were the most efficient and costeffective light source for an algal photobioreactor. They can be effectively utilized for nutrient removal from secondary treated wastewater which helps in improving the overall wastewater quality before it is discharged back into the environment.

Keywords: algal bacterial photobioreactor, domestic wastewater, nutrient removal, led bulbs

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783 Study of Microbial Diversity Associated with Tarballs and Their Exploitation in Crude Oil Degradation

Authors: Varsha Shinde, Belle Damodara Shenoy

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Tarballs are crude oil remnants found in oceans after long term weathering process and are a global concern since several decades as potential marine pollutant. Being complicated in structure microbial remediation of tarballs in natural environment is a slow process. They are rich in high molecular weight alkanes and poly aromatic hydrocarbons which are resistant to microbial attack and other environmental factors, therefore remain in environment for long time. However, it has been found that many bacteria and fungi inhabit on tarballs for nutrients and shelter. Many of them are supposed to be oil degraders, while others are supposed to be getting benefited by byproducts formed during hydrocarbon metabolism. Thus tarballs are forming special interesting ecological niche of microbes. This work aimed to study diversity of bacteria and fungi from tarballs and to see their potential application in crude oil degradation. The samples of tarballs were collected from Betul beach of south Goa (India). Different methods were used to isolate culturable fraction of bacteria and fungi from it. Those were sequenced for 16S rRNA gene and ITS for molecular level identification. The 16S rRNA gene sequence analysis revealed the presence of 13 bacterial genera/clades (Alcanivorax, Brevibacterium, Bacillus, Cellulomonas, Enterobacter, Klebsiella, Marinobacter, Nitratireductor, Pantoea, Pseudomonas, Pseudoxanthomonas, Tistrella and Vibrio), while the ITS sequence analysis placed the fungi in 8 diverse genera/ clades (Aspergillus, Byssochlamys, Monascus, Paecilomyces, Penicillium, Scytalidium/ Xylogone, Talaromyces and Trichoderma). All bacterial isolates were screened for oil degradation capacity. Potential strains were subjected to crude oil degradation experiment for quantification. Results were analyzed by GC-MS-MS.

Keywords: bacteria, biodegradation, crude oil, diversity, fungi, tarballs

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782 Effect of Addition Cinnamon Extract (Cinnamomum burmannii) to Water Content, pH Value, Total Lactid Acid Bacteria Colonies, Antioxidant Activity and Cholesterol Levels of Goat Milk Yoghurt Isolates Dadih (Pediococcus pentosaceus)

Authors: Endang Purwati, Ely Vebriyanti, R. Puji Hartini, Hendri Purwanto

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This study aimed to determine the effect of addition cinnamon extract (Cinnamomum burmannii) in making goat milk yogurt product isolates dadih (Pediococcus pentosaceus) to antioxidant activity and cholesterol levels. The method of research was the experimental method by using a Randomized Block Design (RBD), which consists of 5 treatments with 4 groups as replication. Treatment in this study was used of cinnamon extract as A (0%), B (1%), C (2%), D (3%), E (4%) in a goat’s milk yoghurt. This study was used 4200 ml of Peranakan Etawa goat’s milk and 80 ml of cinnamon extract. The variable analyzed were water content, pH value, total lactic acid bacterial colonies, antioxidant activity and cholesterol levels. The average water content ranged from 81.2-85.56%. Mean pH values rang between 4.74–4.30. Mean total lactic acid bacteria colonies ranged from 3.87 x 10⁸ - 7.95 x 10⁸ CFU/ml. The average of the antioxidant activity ranged between 10.98%-27.88%. Average of cholesterol levels ranged from 14.0 mg/ml–17.5 mg/ml. The results showed that the addition of cinnamon extract in making goat milk yoghurt product isolates dadih (Pediococcus pentosaceus) significantly different (P < 0.05) to water content, pH value, total lactic acid bacterial colonies, antioxidant activity and cholesterol levels. In conclusion, the study shows that using of cinnamon extract 4% is the best in making goat milk yoghurt.

Keywords: antioxidant, cholesterol, cinnamon, Pediococcus pentosaceus, yoghurt

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781 Bacteriological Spectrum and Resistance Patterns of Common Clinical Isolates from Infections in Cancer Patients

Authors: Vivek Bhat, Rohini Kelkar, Sanjay Biswas

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Introduction: Cancer patients are at increased risk of bacterial infections. This may due to the disease process itself, the effect of chemotherapeutic drugs or invasive procedures such as catheterization. A wide variety of bacteria including some emerging pathogens are increasingly being reported from these patients. The incidence of multidrug-resistant organisms particularly in the Gram negative group is also increasing, with higher resistance rates seen to cephalosporins, β-lactam/β-lactam inhibitor combinations, and the carbapenems. This study documents the bacteriological spectrum of infections and their resistance patterns in cancer patients. Methods: This study includes all bacterial isolates recovered from infections cancer patients over a period of 18 months. Samples included Blood cultures, Pus/wound swabs, urine, tissue biopsies, body fluids, catheter tips and respiratory specimens such as sputum and bronchoalveolar lavage (BAL). All samples were processed in the microbiology laboratory as per standard laboratory protocols. Organisms were identified to species level and antimicrobial susceptibility testing was performed manually by the disc diffusion technique or in the Vitek-2 (Biomereux, France) instrument. Interpretations were as per Clinical laboratory Standards Institute (CLSI) guidelines. Results: A total of 1150 bacterial isolates were cultured from 884 test samples during the study period. Of these 227 were Gram-positive and 923 were Gram-negative organisms. Staphylococcus aureus (99 isolates) was the commonest Gram-positive isolate followed by Enterococcus (79) and Gr A Streptococcus (30). Among the Gram negatives, E. coli (304), Pseudomonas aeruginosa (201) and Klebsiella pneumoniae (190) were the most common. Of the Staphylococcus aureus isolates 27.2% were methicillin resistant. Only 5.06% enterococci were vancomycin resistant. High rates of resistance to cefotaxime and ciprofloxacin were seen amongst E. coli (84.8% & 83.55%) and Klebsiella pneumoniae (71 & 62.1%) respectively. Resistance to carbapenems (meropenem) was high at 70% in Acinetobacter spp.; however all isolates were sensitive to colistin. Among the aminoglycosides, amikacin retained good efficacy against Escherichia coli (82.9%) and Pseudomonas aeruginosa (78.1%). Occasional isolates of emerging pathogens such as Chryseobacterium indologens, Roseomonas, and Achromobacter xyloxidans were also recovered. Conclusion: The common infections in cancer patients include respiratory, wound, tract infections and sepsis. The commonest isolates include Staphylococcus aureus, Enterococci, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. There is a high level of resistance to the commonly used antibiotics among Gram-negative organisms.

Keywords: bacteria, resistance, infection, cancer

Procedia PDF Downloads 276
780 Effectiveness of Essential Oils as Inhibitors of Quorum Sensing Activity Using Biomonitor Strain Chromobacterium Violaceum

Authors: Ivana Cabarkapa, Zorica Tomicic, Olivera Duragic

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Antimicrobial resistance represents one of the major challenges facing humanity in the last decades. Increasing antibiotic-resistant pathogens indicates the need for the development of alternative antibacterial drugs and new treatment strategies. One of the innovative emerging treatments in overcoming multidrug-resistant pathogens certainly represents the inhibition anti-quorum sensing system. For most of the food-borne pathogens, the expression of the virulence depends on their capability communication with other members of the population by means of quorum sensing (QS). QS represents a specific way of bacterial intercellular communication, which enabled owing to their ability to detect and to respond to cell population density by gene regulation. QS mechanisms are responsible for controls the pathogenesis, virulence luminescence, motility, sporulation and biofilm formation of many organisms by regulating gene expression. Therefore, research in this field is being an attractive target for the development of new natural antibacterial agents. Anti-QS compounds are known to have the ability to prohibit bacterial pathogenicity. Considering the importance of quorum sensing during bacterial pathogenesis, this research has been focused on evaluation anti - QS properties of four essential oils (EOs) Origanum heracleoticum, Origanum vulgare, Thymus vulgare, and Thymus serpyllum, using biomonitor strain of Chromobacterium violaceum CV026. Tests conducted on Luria Bertani agar supplemented with N hexanol DL homoserine lacton (HHL) 10µl/50ml of agar. The anti-QS potential of the EOs was assayed in a range of concentrations of 200 – 0.39 µl/ml using the disc diffusion method. EOs of Th. vulgaris and T. serpyllum were exhibited anti-QS activity indicated by a non- pigmented ring with a dilution-dependent manner. The lowest dilution of EOs T. vulgaris and T. serpyllum in which they exhibited visually detectable inhibition of violacein synthesis was 6.25 µl/ml for both tested EOs. EOs of O. heracleoticum and O. vulgare were displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by the outer non-pigmented ring, in a concentration-dependent manner. The lowest dilution of EOs of O. heracleoticum and O. vulgare in which exhibited visually detectable inhibition of violacein synthesis was 1.56 and 3.25 µl/ml, respectively. Considering that, the main constituents of the tested EOs represented by monoterpenes (carvacrol, thymol, γ-terpinene, and p-cymene), anti - QS properties of tested EOs can be mainly attributed to their activity. In particular, from the scientific literature, carvacrol and thymol show a sub-inhibitory effect against foodborne pathogens. Previous studies indicated that sub-lethal concentrations of carvacrol reduced the mobility of bacteria due to the ability of interference using QS mechanism between the bacterial cells, and thereby reducing the ability of biofilm formation The precise mechanism by which carvacrol inhibits biofilm formation is still not fully understood. Our results indicated that EOs displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by an outer non- pigmented ring with visually detectable inhibition of violacein. Preliminary results suggest that EOs represent a promising alternative for effective control of the emergence and spread of resistant pathogens.

Keywords: anti-quorum sensing activity, Chromobacterium violaceum, essential oils, violacein

Procedia PDF Downloads 107
779 Probiotic Antibacterial Test of Pediococcus pentosaceus Isolated from Dadih in Inhibiting Periodontitis Bacteria: In Vitro Study on Bacteria Aggregatibacter actinomycetemcomitans

Authors: Nurlaili Syafar Wulan, Almurdi, Suprianto Kosno

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Introduction: Periodontitis defined as an inflammatory disease of teeth supporting tissue with irritation of specific pathogens as the main aetiology. Periodontitis can be cured by giving medical action accompanied by administration of an antibiotic, but the use of antibiotic has a side effect that can cause bacterial resistance. This side effect can be corrected by probiotic, which has antibiotic-like substance but do not have bacterial resistance effect; it makes probiotic became a promising future periodontitis medication. West Sumatran people has their own typical traditional food product made from fermented buffalo’s milk called dadih, and it contained probiotics. Objectives: The aim of this study was to determine the ability of probiotic Pediococcus pentosaceus isolated from dadih in inhibiting the growth of bacteria Aggregatibacter actinomycetemcomitans. Material and Method: This was a true experimental study with post-test and control group design. This study was conducted on 36 samples of 2 treatment groups, the test group with probiotic Pediococcus pentosacesus isolated from dadih and the negative control group with sterile aquadest. The antibacterial effect was tested using the Kirby-Bauer disk diffusion method and calculated by measuring the zone of inhibition on MHA around paper disk using a sliding caliper with 0.5 mm accuracy. Result: The result of bivariate analysis using Independent t-test was p=0.00 where p < 0.05 means that there is a significant difference between the tested group and negative control group. Conclusion: Probiotic Pediococcus pentosaceus isolated from dadih are able to inhibit the growth of Aggregatibacter actinomycetemcomitans.

Keywords: aggregatibacter actinomycetemcomitans, antibacterial activities, periodontitis, probiotic Pediococcus pentosaceus

Procedia PDF Downloads 105
778 Degradation of the Cu-DOM Complex by Bacteria: A Way to Increase Phytoextraction of Copper in a Vineyard Soil

Authors: Justine Garraud, Hervé Capiaux, Cécile Le Guern, Pierre Gaudin, Clémentine Lapie, Samuel Chaffron, Erwan Delage, Thierry Lebeau

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The repeated use of Bordeaux mixture (copper sulphate) and other chemical forms of copper (Cu) has led to its accumulation in wine-growing soils for more than a century, to the point of modifying the ecosystem of these soils. Phytoextraction of copper could progressively reduce the Cu load in these soils, and even to recycle copper (e.g. as a micronutrient in animal nutrition) by cultivating the extracting plants in the inter-row of the vineyards. Soil cleaning up usually requires several years because the chemical speciation of Cu in solution is mainly based on forms complexed with dissolved organic matter (DOM) that are not phytoavailable, unlike the "free" forms (Cu2+). Indeed, more than 98% of Cu in the solution is bound to DOM. The selection and inoculation of invineyardsoils in vineyard soils ofbacteria(bioaugmentation) able to degrade Cu-DOM complexes could increase the phytoavailable pool of Cu2+ in the soil solution (in addition to bacteria which first mobilize Cu in solution from the soil bearing phases) in order to increase phytoextraction performance. In this study, sevenCu-accumulating plants potentially usable in inter-row were tested for their Cu phytoextraction capacity in hydroponics (ray-grass, brown mustard, buckwheat, hemp, sunflower, oats, and chicory). Also, a bacterial consortium was tested: Pseudomonas sp. previously studied for its ability to mobilize Cu through the pyoverdine siderophore (complexing agent) and potentially to degrade Cu-DOM complexes, and a second bacterium (to be selected) able to promote the survival of Pseudomonas sp. following its inoculation in soil. Interaction network method was used based on the notions of co-occurrence and, therefore, of bacterial abundance found in the same soils. Bacteria from the EcoVitiSol project (Alsace, France) were targeted. The final step consisted of incoupling the bacterial consortium with the chosen plant in soil pots. The degradation of Cu-DOMcomplexes is measured on the basis of the absorption index at 254nm, which gives insight on the aromaticity of the DOM. The“free” Cu in solution (from the mobilization of Cu and/or the degradation of Cu-MOD complexes) is assessed by measuring pCu. Eventually, Cu accumulation in plants is measured by ICP-AES. The selection of the plant is currently being finalized. The interaction network method targeted the best positive interactions ofFlavobacterium sp. with Pseudomonassp. These bacteria are both PGPR (plant growth promoting rhizobacteria) with the ability to improve the plant growth and to mobilize Cu from the soil bearing phases (siderophores). Also, these bacteria are known to degrade phenolic groups, which are highly present in DOM. They could therefore contribute to the degradation of DOM-Cu. The results of the upcoming bacteria-plant coupling tests in pots will be also presented.

Keywords: complexes Cu-DOM, bioaugmentation, phytoavailability, phytoextraction

Procedia PDF Downloads 58
777 A Proteomic Approach for Discovery of Microbial Cellulolytic Enzymes

Authors: M. S. Matlala, I. Ignatious

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Environmental sustainability has taken the center stage in human life all over the world. Energy is the most essential component of our life. The conventional sources of energy are non-renewable and have a detrimental environmental impact. Therefore, there is a need to move from conventional to non-conventional renewable energy sources to satisfy the world’s energy demands. The study aimed at screening for microbial cellulolytic enzymes using a proteomic approach. The objectives were to screen for microbial cellulases with high specific activity and separate the cellulolytic enzymes using a combination of zymography and two-dimensional (2-D) gel electrophoresis followed by tryptic digestion, Matrix-assisted Laser Desorption Ionisation-Time of Flight (MALDI-TOF) and bioinformatics analysis. Fungal and bacterial isolates were cultured in M9 minimal and Mandel media for a period of 168 hours at 60°C and 30°C with cellobiose and Avicel as carbon sources. Microbial cells were separated from supernatants through centrifugation, and the crude enzyme from the cultures was used for the determination of cellulase activity, zymography, SDS-PAGE, and two-dimensional gel electrophoresis. Five isolates, with lytic action on carbon sources studied, were a bacterial strain (BARK) and fungal strains (VCFF1, VCFF14, VCFF17, and VCFF18). Peak cellulase production by the selected isolates was found to be 3.8U/ml, 2.09U/ml, 3.38U/ml, 3.18U/ml, and 1.95U/ml, respectively. Two-dimensional gel protein maps resulted in the separation and quantitative expression of different proteins by the microbial isolates. MALDI-TOF analysis and database search showed that the expressed proteins in this study closely relate to different glycoside hydrolases produced by other microbial species with an acceptable confidence level of 100%.

Keywords: cellulases, energy, two-dimensional gel electrophoresis, matrix-assisted laser desorption ionisation-time of flight, MALDI-TOF MS

Procedia PDF Downloads 106
776 A Case Report on Neonatal Conjunctivitis in Pugs

Authors: Maria L. G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Neonatal conjunctivitis, or ophthalmia, is an infection of the conjunctiva or cornea before opening the eyelids. It is believed that immunodeficiency contributes to the development of the condition. This study aims at reporting a case of ophthalmia neonatorum in a dog, in addition to its diagnosis and treatment. A litter of five pug neonates was admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, with complaints of ocular secretion. The neonates were five days old. The clinical examination revealed that three newborns presented swelling in the ocular region and a purulent secretion in the medial corner of the eye that was exerting pressure on the ocular globes, which are compatible with the description of this disease. The diagnosis was made based on the clinical signs and bacterial culture of the secretion, which revealed the presence of bacteria belonging to the genus Staphylococcus sp. The laboratory assays did not reveal any alterations. The treatment was instituted gently, opening the eyelids early and cleaning the purulent ocular secretion with saline solution. An ophthalmic ointment with retinol, amino acids, methionine, and chloramphenicol (Epitezan®) was prescribed four times a day for seven days. Blood plasma (2 mL/100 g) was administered subcutaneously because bacterial infections in neonates may represent a failure in the transference of passive immunity. A more thorough cleaning of the environment was also recommended. Neonatal conjunctivitis has a simple diagnosis and treatment. If not treated early, it can evolve to adherence of the eyelids to the cornea, ulceration, and perforation of the cornea. Therefore, the prognosis is favorable as long as the condition is diagnosed early, and the treatment is instituted quickly.

Keywords: ophthalmia neonatorum, neonatal infection, puppy, newborn

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775 Antibacterial Activity of Copper Nanoparticles on Vancomycin Resistant Staphylococcus Aureus in Vitro and Animal Models

Authors: Sina Gharevali

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Staphylococcus aureus is one of the most important factors for nosocomial infections and infections acquired in a hospital setting role as is. Drug-resistant bacteria methicillin, which in 1961 was reported in many parts of the world, Made the role as the last drug, vancomycin, in the treatment of infections caused by the Staphylococcus aureus chain be taken into consideration. The aim of this study was to evaluate the antimicrobial effects of copper nanoparticles and compared it with antibiotics on Staphylococcus aureus resistant to vancomycin in vitro and animal model. In this study, this test was performed, and the most effective antibiotic for vancomycin-resistant Staphylococcus aureus was determined by disk diffusion method. After various concentrations of copper nanoparticles and antibiotics were prepared and vancomycin resistant Staphylococcus aureus bacteria with serial dilution method for determining antibiotic ciprofloxacin. Minimum Inhibitory Concentration and Minimum Bactericidal Concentrationcopper nanoparticles was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. Then the broth dilution method for the antibiotic ciprofloxacin, nano-particles, and nano-particles of copper and copper-established antibiotic synergy MIC and MBC were obtained. MBC was obtained from the experimental animal model test method, and the results were compared. The results showed that copper nanoparticles compared with the antibiotic ciprofloxacin in vitro and animal model more effective in inhibiting the growth of Staphylococcus aureus resistant to vancomycin and ciprofloxacin and extent of the impact of the Synthetic effect of lower copper nanoparticles. Which can then be used to treat clinical research as a candidate.

Keywords: nanoparticles, copper, staphylococcus, aureus

Procedia PDF Downloads 72
774 Isolation and Characterisation of Novel Environmental Bacteriophages Which Target the Escherichia coli Lamb Outer Membrane Protein

Authors: Ziyue Zeng

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Bacteriophages are viruses which infect bacteria specifically. Over the past decades, phage λ has been extensively studied, especially its interaction with the Escherichia coli LamB (EcLamB) protein receptor. Nonetheless, despite the enormous numbers and near-ubiquity of environmental phages, aside from phage λ, there is a paucity of information on other phages which target EcLamB as a receptor. In this study, to answer the question of whether there are other EcLamB-targeting phages in the natural environment, a simple and convenient method was developed and used for isolating environmental phages which target a particular surface structure of a particular bacterium; in this case, the EcLamB outer membrane protein. From the enrichments with the engineered bacterial hosts, a collection of EcLamB-targeting phages (ΦZZ phages) were easily isolated. Intriguingly, unlike phage λ, an obligate EcLamB-dependent phage in the Siphoviridae family, the newly isolated ΦZZ phages alternatively recognised EcLamB or E. coli OmpC (EcOmpC) as a receptor when infecting E. coli. Furthermore, ΦZZ phages were suggested to represent new species in the Tequatrovirus genus in the Myoviridae family, based on phage morphology and genomic sequences. Most phages are thought to have a narrow host range due to their exquisite specificity in receptor recognition. With the ability to optionally recognise two receptors, ΦZZ phages were considered relatively promiscuous. Via the heterologous expression of EcLamB on the bacterial cell surface, the host range of ΦZZ phages was further extended to three different enterobacterial genera. Besides, an interesting selection of evolved phage mutants with a broader host range was isolated, and the key mutations involved in their evolution to adapt to new hosts were investigated by genomic analysis. Finally, and importantly, two ΦZZ phages were found to be putative generalised transducers, which could be exploited as tools for DNA manipulations.

Keywords: environmental microbiology, phage, microbe-host interactions, microbial ecology

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773 Improved Production, Purification and Characterization of Invertase from Penicillium lilacinum by Shaken Flask Technique of Submerged Fermentation

Authors: Kashif Ahmed

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Recent years researchers have been motivated towards extensive exploring of living organism, which could be utilized effectively in intense industrial conditions. The present study shows enhanced production, purification and characterization of industrial enzyme, invertase (Beta-D-fructofuranosidase) from Penicillium lilacinum. Various agricultural based by-products (cotton stalk, sunflower waste, rice husk, molasses and date syrup) were used as energy source. The highest amount of enzyme (13.05 Units/mL) was produced when the strain was cultured on growth medium containing date syrup as energy source. Yeast extract was used as nitrogen source after 96 h of incubation at incubation temperature of 40º C. Initial pH of medium was 8.0, inoculum size 6x10⁶ conidia and 200 rev/min agitation rate. The enzyme was also purified (7 folds than crude) and characterized. Molecular mass of purified enzyme (65 kDa) was determined by 10 % SDS-PAGE. Lineweaver-Burk Plot was used to determine Kinetic constants (Vmax 178.6 U/mL/min and Km 2.76 mM). Temperature and pH optima were 55º C and 5.5 respectively. MnCl₂ (52.9 %), MgSO₄ (48.9 %), BaCl₂ (24.6 %), MgCl₂ (9.6 %), CoCl₂ (5.7 %) and NaCl (4.2 %) enhanced the relative activity of enzyme and HgCl₂ (-92.8 %), CuSO₄ (-80.2 %) and CuCl₂ (-76.6 %) were proved inhibitors. The strain was showing enzyme activity even at extreme conditions of temperature (up to 60º C) and pH (up to 9), so it can be used in industries.

Keywords: invertase, Penicillium lilacinum, submerged fermentation, industrial enzyme

Procedia PDF Downloads 122
772 Bacterial Interactions of Upper Respiratory Tract Microbiota

Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys

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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.

Keywords: nasal, microbiota, S.aureus, microbioal interaction

Procedia PDF Downloads 76
771 Harnessing the Generation of Ferromagnetic and Silver Nanostructures from Tropical Aquatic Microbial Nanofactories

Authors: Patricia Jayshree Jacob, Mas Jaffri Masarudinb, Mohd Zobir Hussein, Raha Abdul Rahim

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Iron based ferromagnetic nanoparticles (IONP) and silver nanostructures (AgNP) have found a wide range of application in antimicrobial therapy, cell targeting, and environmental applications. As such, the design of well-defined monodisperse IONPs and AgNPs have become an essential tool in nanotechnology. Fabrication of these nanostructures using conventional methods is not environmentally conducive and weigh heavily on energy and outlays. Selected microorganisms possess the innate ability to reduce metallic ions in colloidal aqueous solution to generate nanoparticles. Hence, harnessing this potential is a way forward in constructing microbial nano-factories, capable of churning out high yields of well-defined IONP’s and AgNP's with physicochemical characteristics on par with the best synthetically produced nanostructures. In this paper, we report the isolation and characterization of bacterial strains isolated from the tropical marine and freshwater ecosystems of Malaysia that demonstrated facile and rapid generation of ferromagnetic nanoparticles and silver nanostructures when precursors such as FeCl₃.6H₂O and AgNO₃ were added to the cell-free bacterial lysate in colloidal solution. Characterization of these nanoparticles was carried out using FESEM, UV Spectrophotometer, XRD, DLS and FTIR. This aerobic bioprocess was carried out at ambient temperature and humidity and has the potential to be developed for environmental friendly, cost effective large scale production of IONP’s. A preliminary bioprocess study on the harvesting time, incubation temperature and pH was also carried out to determine pertinent abiotic parameters contributing to the optimal production of these nanostructures.

Keywords: iron oxide nanoparticles, silver nanoparticles, biosynthesis, aquatic bacteria

Procedia PDF Downloads 255
770 The Ability of Consortium Wastewater Protozoan and Bacterial Species to Remove Chemical Oxygen Demand in the Presence of Nanomaterials under Varying pH Conditions

Authors: Anza-Vhudziki Mboyi, Ilunga Kamika, Maggy Momba

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The aim of this study was to ascertain the survival limit and capability of commonly found wastewater protozoan (Aspidisca sp, Trachelophyllum sp, and Peranema sp) and bacterial (Bacillus licheniformis, Brevibacillus laterosporus, and Pseudomonas putida) species to remove COD while exposed to commercial nanomaterials under varying pH conditions. The experimental study was carried out in modified mixed liquor media adjusted to various pH levels (pH 2, 7 and 10), and a comparative study was performed to determine the difference between the cytotoxicity effects of commercial zinc oxide (nZnO) and silver (nAg) nanomaterials (NMs) on the target wastewater microbial communities using standard methods. The selected microbial communities were exposed to lethal concentrations ranging from 0.015 g/L to 40 g/L for nZnO and from 0.015 g/L to 2 g/L for nAg for a period of 5 days of incubation at 30°C (100 r/min). Compared with the absence of NMs in wastewater mixed liquor, the relevant environmental concentration ranging between 10 µg/L and 100 µg/L, for both nZnO and nAg caused no adverse effects, but the presence of 20 g of nZnO/L and 0.65 g of nAg/L significantly inhibited microbial growth. Statistical evidence showed that nAg was significantly more toxic compared to nZnO, but there was an insignificant difference in toxicity between microbial communities and pH variations. A significant decrease in the removal of COD by microbial populations was observed in the presence of NMs with a moderate correlation of r = 0.3 to r = 0.7 at all pH levels. It was evident that there was a physical interaction between commercial NMs and target wastewater microbial communities; although not quantitatively assessed, cell morphology and cell death were observed. Such phenomena suggest the high resilience of the microbial community, but it is the accumulation of NMs that will have adverse effects on the performance in terms of COD removal.

Keywords: bacteria, biological treatment, chemical oxygen demand (COD) and nanomaterials, consortium, pH, protozoan

Procedia PDF Downloads 272
769 A Viable Approach for Biological Detoxification of Non Edible Oil Seed Cakes and Their Utilization in Food Production Using Aspergillus Niger

Authors: Kshitij Bhardwaj, R.K. Trivedi, Shipra Dixit

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We used biological detoxification method that converts toxic residue waste of Jatropha curcas oil seeds (non edible oil seed) into industrial bio-products and animal feed material. Present study describes the complete degradation of phorbol esters by Aspergillus Niger strain during solid state fermentation (SSF) of deoiled Jatropha curcas seed cake. Phorbol esters were completely degraded in 15 days under the optimized SSF conditions viz deoiled cake 5.0 gm moistened with 5.0 ml distilled water; inoculum 2 ml of overnight grown Aspergillus niger; incubated at 30◦ C, pH 7.0. This method simultaneously induces the production of Protease enzyme by Aspergillus Niger which has high potential to be used in feedstuffs .The maximum Protease activities obtained were 709.16 mg/ml in Jatropha curcas oil seed cake. The protein isolate had small amounts of phorbol esters, phytic acid, and saponin without any lectin. Its minimum and maximum solubility were at pH 4.0&12.0. Water and oil binding capacities were 3.22 g water/g protein and 1.86 ml oil/g protein respectively.Emulsion activity showed high values in a range of basic pH. We concluded that Jatropha Curcas seed cake has a potential to be used as a novel source of functional protein for food or feed applications.

Keywords: solid state fermentation, Jatropha curcas, oil seed cake, phorbol ester

Procedia PDF Downloads 455
768 Bacteriological Quality and Physicochemical Water Beaches of the City of Annaba (Mediterranean Sea)

Authors: Wahiba Boudraa, Farah Chettibbi, Meriem Aberkane, Fatma Djamaa, Moussa Houhamdi

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The intensity of human activities in regions surrounding the Mediterranean Sea always has a strong long-term environmental impact resulting in coastal and marine degradation, as well as an aggravated risk of more serious damage. The available data on water quality show that most water resources in Algeria are polluted by uncontrolled discharges from municipal sewage and untreated industrial effluents. Annaba is a coastal town in Algeria; The Gulf of Annaba, responds to these changes as it receives the continental inputs and urban waste, industrial without prior treatment of a highly industrialized and urbanized city, subject to the same environmental problems that know the rest of the Algerian coast. In later year, the beaches of bacterial enumeration process waters showed relatively high levels of bacterial indicators of fecal contamination (group D streptococci, total and fecal coliforms), which reflect the risks to people attending these beaches. During the twelve months of our study, we isolated from three beaches in the city of Annaba (St. Cloud, El-Kettara, and Djenane El Bey) a number of pathogenic microorganisms considered, namely: Salmonella, Aeromonas, Citrobacter, Yersinia, Enterococcus, and E.coli. The microbial count revealed elevated levels of coliform bacteria, fecal coliforms and fecal streptococci quite high especially in urban beaches (St. Cloud and El-Kettara). They are widely popular during the summer by many vacationers. For the physico-chemical parameters, there exist some weak values which increase during the pluvial period, hivernal and festival saison. These values remain, nevertheless, weak to be able to cause an organic or metallic pollution.

Keywords: quality microbiology, pollution of water, fecal contamination, physico-chemistry, beaches of Annaba city, Algeria.

Procedia PDF Downloads 318
767 Analysis of Chemical Composition of Raw Milk in Some Farms Animals in El Khoms, Libya

Authors: Dukali Abujnah

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This study was undertaken to knowledge the chemical composition of some farm animals (cows ) by using ultrasonic milk analyzer. This method was used first time in our country in the field of milk hygiene to determine and comparison the composition percent of raw milk for farm animals as attempt to protect the health of consumers 240 raw milk samples for each species of animals included in this study for two different season of year first (Novembe 2014 - January 2015) , second from (May – Novembe2015) .The chemical analysis. In this way, a range of building suitable to put in the Libyan market product structures, and milk, for use as a new product or exposure to treatment such as pasteurized milk and long-life may be wider study project..Controls have been implemented in operating in Libya, companies which are part of the business created by the Libyan state. It was created only after the companies that made many of the studies and research carried out on areas of Libya which it was possible to intervene. Although troops spent in recent years, companies have not yet found their balance and milk production has not yet come to those set forth in the action plans, due to problems of various types that will be discussed in this study. We believe that these data have been taken in four years if analyzed properly, can provide more accurate information to farmers in their companies want to take dairy cattle breeding. The total bacterial count was 76 and 38 cfu/ml for locally produced and imported UHT milk respectively. Coliforms were negative for all examined locally produce and imported samples. E. coli were not isolated from any of the examined UHT samples. Aerobic spore-formers count mean count was 49 and 27 cfu/ml for both examined samples respectively. Obtained results show that most of the locally produced UHT milk does not satisfy the suggested Libyan Standards for UHT milk for fat% as well as total bacterial count/ml. Economical and public health importance of the obtained results were discussed, and control measures for improving the quality of UHT locally made milk were also discussed.

Keywords: locally, UHT milks, farms animals, raw milk analysis milk

Procedia PDF Downloads 262