Search results for: sulphur oxidizing bacteria
Commenced in January 2007
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Edition: International
Paper Count: 1691

Search results for: sulphur oxidizing bacteria

1031 Assessment on Rumen Microbial Diversity of Bali Cattle Using 16S rRNA Sequencing

Authors: Asmuddin Natsir, A. Mujnisa, Syahriani Syahrir, Marhamah Nadir, Nurul Purnomo

Abstract:

Bacteria, protozoa, Archaea, and fungi are the dominant microorganisms found in the rumen ecosystem that has an important role in converting feed ingredients into components that can be digested and utilized by the livestock host. This study was conducted to assess the diversity of rumen bacteria of bali cattle raised under traditional farming condition. Three adult bali cattle were used in this experiment. The rumen fluid samples from the three experimental animals were obtained by the Stomach Tube method before the morning feeding. The results of study indicated that the Illumina sequencing was successful in identifying 301,589 sequences, averaging 100,533 sequences, from three rumen fluid samples of three cattle. Furthermore, based on the SILVA taxonomic database, there were 19 kinds of phyla that had been successfully identified. Of the 19 phyla, there were only two dominant groups across the three samples, namely Bacteroidetes and Firmicutes, with an average percentage of 83.68% and 13.43%, respectively. Other groups such as Synergistetes, Spirochaetae, Planctomycetes can also be identified but in relatively small percentage. At the genus level, there were 157 sequences obtained from all three samples. Of this number, the most dominant group was Prevotella 1 with a percentage of 71.82% followed by 6.94% of Christencenellaceae R-7 group. Other groups such as Prevotellaceae UCG-001, Ruminococcaceae NK4A214 group, Sphaerochaeta, Ruminococcus 2, Rikenellaceae RC9 gut group, Quinella were also identified but with very low percentages. The sequencing results were able to detect the presence of 3.06% and 3.92% respectively for uncultured rumen bacterium and uncultured bacterium. In conclusion, the results of this experiment can provide an opportunity for a better understanding of the rumen bacterial diversity of the bali cattle raised under traditional farming condition and insight regarding the uncultured rumen bacterium and uncultured bacterium that need to be further explored.

Keywords: 16S rRNA sequencing, bali cattle, rumen microbial diversity, uncultured rumen bacterium

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1030 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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1029 Wind Direction and Its Linkage with Vibrio cholerae Dissemination

Authors: Shlomit Paz, Meir Broza

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Cholera is an acute intestinal infection caused by ingestion of food or water contaminated with the bacterium Vibrio cholerae. It has a short incubation period and produces an enterotoxin that causes copious, painless, watery diarrhoea that can quickly lead to severe dehydration and death if treatment is not promptly given. In an epidemic, the source of the contamination is usually the feces of an infected person. The disease can spread rapidly in areas with poor treatment of sewage and drinking water. Cholera remains a global threat and is one of the key indicators of social development. An estimated 3-5 million cases and over 100,000 deaths occur each year around the world. The relevance of climatic events as causative factors for cholera epidemics is well known. However, the examination of the involvement of winds in intra-continental disease distribution is new. The study explore the hypothesis that the spreading of cholera epidemics may be related to the dominant wind direction over land by presenting the influence of the wind direction on windborn dissemination by flying insects, which may serve as vectors. Chironomids ("non-biting midges“) exist in the majority of freshwater aquatic habitats, especially in estuarine and organic-rich water bodies typical to Vibrio cholerae. Chironomid adults emerge into the air for mating and dispersion. They are highly mobile, huge in number and found frequently in the air at various elevations. The huge number of chironomid egg masses attached to hard substrate on the water surface, serve as a reservoir for the free-living Vibrio bacteria. Both male and female, while emerging from the water, may carry the cholera bacteria. In experimental simulation, it was demonstrated that the cholera-bearing adult midges are carried by the wind, and transmit the bacteria from one body of water to another. In our previous study, the geographic diffusions of three cholera outbreaks were examined through their linkage with the wind direction: a) the progress of Vibrio cholerae O1 biotype El Tor in Africa during 1970–1971 and b) again in 2005–2006; and c) the rapid spread of Vibrio cholerae O139 over India during 1992–1993. Using data and map of cholera dissemination (WHO database) and mean monthly SLP and geopotential data (NOAA NCEP-NCAR database), analysis of air pressure data at sea level and at several altitudes over Africa, India and Bangladesh show a correspondence between the dominant wind direction and the intra-continental spread of cholera. The results support the hypothesis that aeroplankton (the tiny life forms that float in the air and that may be caught and carried upward by the wind, landing far from their origin) carry the cholera bacteria from one body of water to an adjacent one. In addition to these findings, the current follow-up study will present new results regarding the possible involvement of winds in the spreading of cholera in recent outbreaks (2010-2013). The findings may improve the understanding of how climatic factors are involved in the rapid distribution of new strains throughout a vast continental area. Awareness of the aerial transfer of Vibrio cholerae may assist health authorities by improving the prediction of the disease’s geographic dissemination.

Keywords: cholera, Vibrio cholerae, wind direction, Vibrio cholerae dissemination

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1028 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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1027 The microbial evaluation of cow raw milk used in private dairy factories in of Zawia city, Libya

Authors: Obied A. Alwan, Elgerbi, M. Ali

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This study was conducted on the cow milk which is used in the local milk factories of Zawia. This was completely random sampling the unscheduled samples. The microbiologic result have approved that the count of bacteria and the count of E.Coli are very high and all the manufacturing places which were included in the study have lacked the health conditions.

Keywords: raw milk, dairy factories, Libya, microbiologic

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1026 Diversity of Culturable Forms of Microorganisms in Soils with Long-term Exposure to Petroleum Hydrocarbons and Prospects for Bioremediation

Authors: Yessentayeva K. Y., Berzhanova R. Z., Mukasheva T. D.

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The purpose of this study was to study the microbial diversity of soils with long-standing hydrocarbon pollution in the S. Balgimbayev field (Kazakhstan), where the transformation of meadow coastal soils technogenic solonchak soils, as well as the assessment of the degradation potential of microorganisms perspective for the use for bioremediation. In the present work autochthonous microorganisms of the surface horizon of soils were investigated. In samples with a low degree of pollution the number of microorganisms, was comparable to the number in the uncontaminated soil and was 103 - 104 CFU/g. and one and two orders of magnitude lower in samples with high oil content. A collection of microorganisms was created using different culture media, which made it possible to isolate isolates that play a key role in different successional stages of biodegradation of petroleum hydrocarbons. The collection included the main bacterial filiiments, Protobacteria, Firmicutes, Actinobacteria and Bacteroidetes. Mycelial fungi andyeast-like fungwere assigned to the Ascomycota division. Studies showed that the percentage of isolates capable of growth in hydrocarbons varied. More than 50 % of the isolates grew on crude oil, a low percentage of less than 10 % of the isolates grew on an anthracene, phenanthrene and naphthalene, more than 20 % of the isolates belonging to different genera Pseudomonas, Bacillus, Rhodococcus, Achromobacter, Gordonia, Microbacterium, and Trichosporon, characterized the growth on two or three different hydrocarbons. The ability to grow using all hydrocarbons, associated with the synthesis of biosurfactants, was detected only in a few isolates.

Keywords: oil, soil, number of bioremediation, biodegradation, microorganisms, hydrocarbons – oxidizing microorganisms

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1025 Endemic Asteraceae from Mauritius Islands as Potential Phytomedicines

Authors: S.Kauroo, J. Govinden Soulange, D. Marie

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Psiadia species from the Asteraceae are traditionally used in the folk medicine of Mauritius to treat cutaneous and bronchial infections. The present study aimed at validating the phytomedicinal properties of the selected species from the Asteraceae family, namely Psiadia arguta, Psiadia viscosa, Psiadia lithospermifolia, and Distephanus populifolius. Dried hexane, ethyl acetate, and methanol leaf extracts were studied for their antioxidant properties using the DPPH (1, 1-diphenyl-2-picryl-hydrazyl), FRAP (Ferric Reducing Ability of Plasma), and Deoxyribose assays. Antibacterial activity against human pathogenic bacteria namely Escherichia coli (ATCC 27853), Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Klebsiella pneumonia (ATCC27853), Pseudomonas aeruginosa (ATCC 27853), and Bacillus cereus (ATCC 11778) was measured using the broth microdilution assay. Qualitative phytochemical screening using standard methods revealed the presence of coumarins, tannins, leucoanthocyanins, and steroids in all the tested extracts. The measured phenolics level of the selected plant extracts varied from 24.0 to 231.6 mg GAE/g with the maximum level in methanol extracts in all four species. The highest flavonoids and proanthocyanidins content was noted in Psiadia arguta methanolic extracts with 65.7±1.8 mg QE/g and 5.1±0.0 mg CAT/g dry weight (DW) extract, respectively. The maximum free radical scavenging activity was measured in Psiadia arguta methanol and ethyl acetate extracts with IC50 11.3±0.2 and 11.6± 0.2 µg/mL, respectively and followed by Distephanus populifolius methanol extracts with an IC50 of 11.3± 0.8 µg/mL. The maximum ferric reducing antioxidant potential was noted in Psiadia lithospermifolia methanol extracts with a FRAP value of 18.8 ± 0.4 µmol Fe2+/L/g DW. The antioxidant capacity based on DPPH and Deoxyribose values were negatively related to total phenolics, flavonoid and proanthocyanidins content while the ferric reducing antioxidant potential were strongly correlated to total phenolics, flavonoid and proanthocyanidins content. All four species exhibited antimicrobial activity against the tested bacteria (both Gram-negative and Gram-positive). Interestingly, the hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia were more active than the control antibiotic Chloramphenicol. The Minimum inhibitory concentration (MIC) values for hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia against the tested bacteria ranged from (62.5 to 500 µg/ml). These findings validate the use of these tested Asteraceae in the traditional medicine of Mauritius and also highlight their pharmaceutical potential as prospective phytomedicines.

Keywords: antibacterial, antioxidant, DPPH, flavonoids, FRAP, Psiadia spp

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1024 Assessment of Air Quality Status Using Pollution Indicators in Industrial Zone of Brega City

Authors: Tawfig Falani, Abdulalaziz Saleh

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Air pollution has become a major environmental issue with definitive repercussions on human health. Global concerns have been raised about the health effects of deteriorating air quality due mainly to widespread industrialization and urbanization. To assess the quality of air in Brega, air quality indicators were calculated using the U.S. Environmental Protection Agency procedure. Air quality was monitored from 01/10/2019 to 28/02/2021 with a daily average measuring six pollutants of particulate matter <2.5µm (PM2.5), and <10µm (PM₁₀), sulfur dioxide (SO₂), nitrogen dioxide (NO₂), ozone (O₃), and carbon monoxide (CO). The result indicated that air pollution at general air quality monitoring sites for sulphur dioxide, carbon monoxide, PM₁₀ and PM2.5 and nitrogen dioxide are always within the permissible limit. Referring to a monthly average of Pollutants in the Brega Industrial area, all months were out of AQG limit for NO₂, and the same with O₃ except for two months. For PM2.5 and PM₁₀ 7, 5 out of 17 months were out of limits, respectively. Relative AQI for ozone is found in the range of moderate category of general air pollution, and the worst month was Nov. 2020, which was marked as Very Unhealthy category, then the next two months (Dec. 2020 and Jan. 2021 ) were Unhealthy categories. It's the first time that we have used the AQI in SOC, and not usually used in Libya to identify the quality of air pollution. So, I think it will be useful if AQI is used as guidance for specified air pollution. That dictate putting monitoring stations beside any industrial activity that has emissions of the six major air pollutants.

Keywords: air quality, air pollutants, air quality index (AQI), particulate matter

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1023 Comparative Analysis of Biodegradation on Polythene and Plastics Buried in Fadama Soil Amended With Organic and Inorganic Fertilizer

Authors: Baba John, Abdullahi Mohammed

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The aim of this research is to compare the analysis of biodegradation on polythene and plastics buried in fadama soil amended with Organic and Inorganic fertilizer. Physico- chemical properties of the samples were determined. Bacteria and Fungi implicated in the biodegradation were identified and enumerated. Physico- chemical properties before the analysis indicated pH range of the samples from 4.28 – 5.80 , While the percentage Organic carbon and Organic matter was highest in cow dung samples with 3.89% and 6.69% respectively. The total Nitrogen percentage was recorded to be highest in Chicken dropping (0.68), while the availability of Phosphorus (P), Sodium (Na), Pottasium (K), and Magnessium (mg) was recorded to be highest in F – soil (Control), with values to be 37ppm, 1.63 Cmolkg-1, 0.35 Cmolkg-1 and 1.18 Cmolkg-1 respectively, except for calcium which was recorded to be highest in Cow dung (5.80 Cmolkg-1). However, physico – chemical properties of the samples after analysis indicated pH range of 4.6 – 5.80, Percentage Organic carbon and Organic matter was highest in Fadama soil mixed with fertilizer, having 0.7% and 1.2% respectively. Total Percentage Nitrogen content was found to be highest (0.56) in Fadama soil mixed with poultry dropping. Availability of Sodium (Na), Pottasium (K), and Calcium (Ca) was recorded to be highest in Fadama Soil mixed with Cow dung with values to be 0.64 Cmolkg-1, 2.07 Cmolkg-1 and 3.36 Cmolkg-1 respectively. The percentage weight loss of polythene and plastic bags after nine months in fadama soil mixed with poultry dropping was 11.9% for polythene and 6.0% for plastics. Weight loss in fadama soil mixed with cow dung was 18.1% for polythene and 4.7% for plastics. Weight loss of polythene and plastic in fadama soil mixed with fertilizer (NPK) was 7.4% for polythene and 3.3% for plastics. While, the percentage weight loss of polythene and plastics after nine months of burial in fadama soil (control) was 3.5% and 0.0% respectively. The bacteria species isolated from Fadama soil, organic and inorganic fertilizers before amendments include: S. aureus, Micrococcus sp, Streptococcus. pyogenes, Psuedomonas aeruginosa Bacillus subtilis and Bacillus cereus. The fungi species include: Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Fusarium sp, Mucor sp Penicillium sp and Candida sp. The bacteria species isolated and characterized after nine months of seeding include: S. aureus, Micrococcus sp, S. pyogenes, P. aeruginosa and B. subtilis. The fungi species are: A. niger A. flavus, A. fumigatus, Mucor sp, Penicillium sp and Fusarium sp. The result of this study indicated that plastic materials can be degraded in the fadama soil irrespective of whether the soil is amended or not. The Period of composting also has a significant impact on the rate at which polythene and plastics are degraded.

Keywords: Fadama, fertilizer, plastic and polythene, biodegradation

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1022 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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1021 Molecular Characterization of Listeria monocytogenes from Fresh Fish and Fish Products

Authors: Beata Lachtara, Renata Szewczyk, Katarzyna Bielinska, Kinga Wieczorek, Jacek Osek

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Listeria monocytogenes is an important human and animal pathogen that causes foodborne outbreaks. The bacteria may be present in different types of food: cheese, raw vegetables, sliced meat products and vacuum-packed sausages, poultry, meat, fish. The most common method, which has been used for the investigation of genetic diversity of L. monocytogenes, is PFGE. This technique is reliable and reproducible and established as gold standard for typing of L. monocytogenes. The aim of the study was characterization by molecular serotyping and PFGE analysis of L. monocytogenes strains isolated from fresh fish and fish products in Poland. A total of 301 samples, including fresh fish (n = 129) and fish products (n = 172) were, collected between January 2014 and March 2016. The bacteria were detected using the ISO 11290-1 standard method. Molecular serotyping was performed with PCR. The isolates were tested with the PFGE method according to the protocol developed by the European Union Reference Laboratory for L. monocytogenes with some modifications. Based on the PFGE profiles, two dendrograms were generated for strains digested separately with two restriction enzymes: AscI and ApaI. Analysis of the fingerprint profiles was performed using Bionumerics software version 6.6 (Applied Maths, Belgium). The 95% of similarity was applied to differentiate the PFGE pulsotypes. The study revealed that 57 of 301 (18.9%) samples were positive for L. monocytogenes. The bacteria were identified in 29 (50.9%) ready-to-eat fish products and in 28 (49.1%) fresh fish. It was found that 40 (70.2%) strains were of serotype 1/2a, 14 (24.6%) 1/2b, two (4.3%) 4b and one (1.8%) 1/2c. Serotypes 1/2a, 1/2b, and 4b were presented with the same frequency in both categories of food, whereas serotype 1/2c was detected only in fresh fish. The PFGE analysis with AscI demonstrated 43 different pulsotypes; among them 33 (76.7%) were represented by only one strain. The remaining 10 profiles contained more than one isolate. Among them 8 pulsotypes comprised of two L. monocytogenes isolates, one profile of three isolates and one restriction type of 5 strains. In case of ApaI typing, the PFGE analysis showed 27 different pulsotypes including 17 (63.0%) types represented by only one strain. Ten (37.0%) clusters contained more than one strain among which four profiles covered two strains; three had three isolates, one with five strains, one with eight strains and one with ten isolates. It was observed that the isolates assigned to the same PFGE type were usually of the same serotype (1/2a or 1/2b). The majority of the clusters had strains of both sources (fresh fish and fish products) isolated at different time. Most of the strains grouped in one cluster of the AscI restriction was assigned to the same groups in ApaI investigation. In conclusion, PFGE used in the study showed a high genetic diversity among L. monocytogenes. The strains were grouped into varied clonal clusters, which may suggest different sources of contamination. The results demonstrated that 1/2a serotype was the most common among isolates from fresh fish and fish products in Poland.

Keywords: Listeria monocytogenes, molecular characteristic, PFGE, serotyping

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1020 Application of Bacteriophages as Natural Antibiotics in Aquaculture

Authors: Chamilani Nikapitiya, Mahanama De Zoysa, Jehee Lee

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Most of the bacterial diseases are associated with high mortalities in aquaculture species and causing huge economic losses. Different approaches have been taken to prevent or control of bacterial diseases including use of vaccines, probiotics, chemotherapy, water quality management, etc. Antibiotics are widely applying as chemotherapy to control bacterial diseases, however, it has been shown that frequent use of antibiotics is favored to develop multi-drug resistance bacteria. Therefore, phages and phage encoded lytic proteins are known to be one of the most promising alternatives for antibiotics to avoid the emergence of antibiotic-resistant bacteria. We isolated and characterized the two lytic phages, namely pAh-1 and pAs-1 against pathogenic Aeromonas hydrophila and Aeromonas salmonicida, respectively. Morphological characteristics were analyzed by Transmission electron microscopy (TEM) and host strain specificities were tested with Aeromonas and other closely related bacterial strains. TEM analysis revealed that both pAh-1 and pAsm-1 are composed of an icosahedral head and a segmented tail, and we suggest that, they are new members of Myoviridae family. Genome sizes of isolated phages were estimated by restriction enzyme digestion of genomic DNA using selected endonucleases followed by agarose gel electrophoresis. Estimated genome size of pAh-1 and pAs-1 were approximately 64 Kbp and 120 Kbp, respectively. Both pAh-1 and pAs-1 have shown narrow host specificity. Moreover, protective effects of phage therapy against fish pathogenic A. hydrophila were investigated in zebrafish model. The survival rate was 40% higher when zebrafish received intra-peritoneal injection (i.p.) of pAh-1 were simultaneously challenge A. hydrophila (2 x 106 CFU/fish) compared to that without phage treatment. Overall results suggest that both pAh-1 and pAs-1 can be used as a potential phage therapy to control Aeromonas infections in aquaculture.

Keywords: Aeromonas infections, antibiotic resistance, bacteriophage, bio-control, lytic phage

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1019 Antimicrobial Activity of Eucalyptus globulus Essential Oil: Disc Diffusion versus Vapour Diffusion Methods

Authors: Boukhatem Mohamed Nadjib, Ferhat Mohamed Amine

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Essential Oils (EO) produced by medicinal plants have been traditionally used for respiratory tract infections and are used nowadays as ethical medicines for colds. The aim of this study was to test the efficacy of the Algerian EGEO against some respiratory tract pathogens by disc diffusion and vapour diffusion methods at different concentrations. The chemical composition of the EGEO was analysed by Gas Chromatography-Mass Spectrometry. Fresh leaves of E. globulus on steam distillation yielded 0.96% (v/w) of essential oil whereas the analysis resulted in the identification of a total of 11 constituents, 1.8 cineole (85.8%), α-pinene (7.2%) and β-myrcene (1.5%) being the main components. By disc diffusion method, EGEO showed potent antimicrobial activity against Gram-positive more than Gram-negative bacteria. The Diameter of Inhibition Zone (DIZ) varied from 69 mm to 75 mm for Staphylococcus aureus and Bacillus subtilis (Gram +) and from 13 to 42 mm for Enterobacter sp and Escherichia coli (Gram-), respectively. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial activity was observed in the vapour phase at lower concentrations. A. baumanii and Klebsiella pneumoniae were the most susceptible strains to the oil vapour with DIZ varied from 38 to 42 mm. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. Else, the DIZ increased with increase in the concentration of the oil. There is growing evidence that EGEO in the vapour phase are effective antibacterial systems and appears worthy to be considered for practical uses in the treatment or prevention of patients with respiratory tract infections or as air decontaminants in the hospital. The present study indicates that EGEO has considerable antimicrobial activity, deserving further investigation for clinical applications.

Keywords: eucalyptus globulus, essential oils, respiratory tract pathogens, antimicrobial activity, vapour phase

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1018 Cytotoxic, Antimicrobial and Antiviral Activities of Acovenoside A: A Cardenolide Isolated from an Egyptian Cultivar of Acokanthera spectabilis Leaves

Authors: Howaida I. Abd-Alla, Amal Z. Hassan, Maha Soltan, Atef G. Hanna, Mounir M. El-Safty

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Acokanthera oblongifolia (Apocynaceae) is used for treatment of several infection diseases and is a well-known cardiac glycoside-containing plant. The infusion of their leaves is gargled to treat tonsillitis and is used medicinally to treat snakebites. The total cardiac glycosides content in the leaves was determined by referring to gitoxigenin as a reference compound. Two triterpenes, lup-20(29)-en-3β-ol (1) and oleanolic acid (2); two cardenolides, acovenoside A (3) and acobioside A (4) were isolated from the ethyl acetate extract. Their structures were determined on the basis of spectral analysis. Major constituents isolated from this species were evaluated for cytotoxicity against normal lung cell line (Wi38) and antimicrobial activities against Gram-positive (two strains) and Gram-negative bacteria (four strains), yeast-like fungi (two strains) and fungi (five strains). The minimum inhibitory concentration (MIC) of the compounds was determined using broth microdilution method. Their viral inhibitory effects against avian influenza virus type A (AI-H5N1) and Newcastle disease virus (NDV) in specific pathogen free (SPF) embryonated chicken eggs (ECE), chicken embryo fibroblasts (CEF) and Vero cells were evaluated. The cardenolide (3) showed viral inhibitory effects against AI-H5N1 and NDV in SPF ECE. The two cardenolides isolated have shown potent cytotoxicity against Vero cells. Compound (3) showed potent anti-Gram-negative bacteria activity. These results suggested that acovenoside A might be promising for future antiviral and antimicrobial drug design.

Keywords: Acokanthera, AI-H5N1, Cardenolides, NDV, SPF-ECE, VERO, Wi38 , Microbe

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1017 Increased Seedling Vigor Through Phytohomeopathy

Authors: Jasper Jose Zanco

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Plants are affected by substances diluted below certain limits. In seeds subjected to ultra-high dilutions (UHD), according to phytohomeopathic methods, it is possible to reduce the concentrations to infinitesimal levels and the effects persist. This research aimed to test different potencies of UHD to modify the vigor of Eruca versicaria (L) Cav. seedlings. The research was carried out at the Plant Production Laboratory of UNISUL University in Santa Catarina, Brazil. Eight UHD treatments were tested, four drops for every 30 mL of distilled water: Control (70% alcohol - A70); Sulphur (S9), Acidum fluoridricum (A30), Calcarea carbonica (C200), Graphies naturalis (G200), Kali carbonicum (K100) Belladonna (B12), diluted and succussed in Hahnemannian centesimal standards. Succussion is a standard pharmaceutical method found in worldwide pharmaceuticals. The statistical design consisted of 50 seeds every 4 replicates per treatment, completely randomized, followed by ANOVA and Tukey's test. Succussion may integrate the high dilution of water treatments, even after successive dilutions, and the product of this process acts through physical-chemical and bioelectric stimuli, causing physiological responses at the cellular level, such as the activation of antioxidant systems, increased resistance to environmental stress or growth modulation. According to some researchers, these responses could be mediated by genetic expression changes or the plants' cellular signaling systems. The results showed significant differences between the control (A70) and the other treatments. Conductivity measurements were made in the seed germination water and impedance; the seedlings were measured for dry weight and total area. The highest conductivity occurred in the control treatment (27.8 μS/cm) and the lowest in K100 (21.3 μS/cm). After germination, on germitest paper, A70 was significantly different from G200 (<1%) and S9 (5%). Both homeopathies differed from the other treatments, with S9 obtaining the best germination (87.1%) and vigor index (IV=7.98) in relation to the other treatments. The control, A70, presented the lowest germination (63.9%) and vigor (IV=4.93).

Keywords: ultra high dilution, impedance, condutivity, eruca versicaria

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1016 Ointment of Rosella Flower Petals Extract (Hibiscus sabdariffa): Pharmaceutical Preparations Formulation Development of Herbs for Antibacterial S. aureus

Authors: Muslihatus Syarifah

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Introduction: Rosella flower petals can be used as an antibacterial because it contains alkaloids, flavonoids, phenolics, and terpenoids) for the . Bacteria activity is S. aureus can cause skin infections and pengobatanya most appropriate use of topical preparations. Ointment is a topical preparation comprising the active substance and ointment base. Not all the base matches the active substances or any type of disease. In this study using flavonoid active substances contained in rosella flower petals (Hibiscus sabdariffa) to be made ointment by testing a variety of different bases in order to obtain a suitable basis for the formulation of ointment extract rosella flower petals. Methods: Experimental research with research methods Post test control group design using the ointment is hydrocarbon sample, absorption, leached water and dissolved water. Then tested for bacteria S. aureus with different concentrations of 1%, 2%, 4%, 8%, 16, 32%. Data were analyzed using One Way ANOVA followed by Post Hoc test. Results: Ointment with a hydrocarbon base, absorption, leached water and dissolved water having no change in physical properties during storage. Base affect the physical properties of an ointment that adhesion, dispersive power and pH. The physical properties of the ointment with different concentrations produce different physical properties including adhesion, dispersive power and pH. The higher the concentration the higher dispersive power, but the smaller the adhesion and pH. Conclusion: Differences bases, storage time, the concentration of the extract can affect the physical properties of the ointment. Concentration of extract in the ointment extract rosella flower petals is 32%.

Keywords: rosella, physical properties, ointments, antibacterial

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1015 The Study of Effective Microorganism's Biopreperation for Wastewater Treatment

Authors: Batsukh Chultem, Oyunbileg Natsagdorj, Namsrai Steyrmunkh

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Many industries, tourist camps and houses, discharge aqueous effluents containing relatively high levels of heavy metals, harmful organic compounds water. Untreated effluent from these manufacturing processes has an adverse impact on the environment. A specific problem associated with waste water in the environment is accumulation in the food chain and persistence in the environment. The screening of microorganisms resistant to pollution and able to detoxification them is essential for the development of clean-up technologies. The purpose of this study is to use advanced microbiological technology products for oxidizing organic and heavy metals pollutants as a biological treatment, to reduce water pollution, which arise as a result of waste water due to day-to-day operations of industries and houses of Ulaanbaatar city and tourist camps located around the lake Hovsgol, in Hovsgol province of Mongolia. By comparing the results from tests of effective microorganism’s bio-preparation treated sewage samples and not treated sewage samples shows that the treated sewage samples pollution decreased defending on treatment period and ratio. Treated water analyses show that: the suspended solids 352 mg/l, pH 5.85-7.95, ammonium nitrate 81.25-221.2 mg NH₄/l, nitrite 0.088-0.227 mg NO₂/l, nitrate 8.5-11.5 mg NO₃/l, and orthophosphate 1.06-15.46 mg PO₄/l. Also, heavy metals were decreased and microbiological test results defined parameters, respectively show the waste water pollution was reduced.

Keywords: effective microorganims, environment, pollution, treatment

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1014 H₆P₂W₁₈O₆₂.14H₂O Catalyzed Synthesis of α-Aminophosphonates from Amino Acids Esters

Authors: Sarra Boughaba

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α-aminophosphonates have found a wide range of applications in organic and medicinal chemistry; they are considered as pharmacological agents, anti-inflammatory antitumor agents, and antibiotics. A number of procedures have been developed for their synthesis. However, many of these methods suffer from some disadvantages such as long reaction times, environmental pollution, utilization of organic solvents, and expensive catalysts. In the past few years, heteropolyacids have received great attention as environmentally benign catalysts for organic synthetic processes, they possess unique physicochemical properties, such as super-acidity, high thermal and chemical stability, ability to accept and release electrons and high proton mobility, and the possibility of varying their acidity and oxidizing potential. In this context, an efficient and eco-friendly protocol has been described for the synthesis of α-aminophosphonates via one pot, three component reaction catalyzed by H₆P₂W₁₈O₆₂.14H₂O as reusable catalyst, by condensation of amino acids esters, various aromatic aldehydes and triethylphosphite under solvent-free conditions, the corresponding α-aminophosphonates were formed in good yields as racemic or diastereomericmixture. All the new products were systematically characterized by IR, MS, and ¹H, ¹³C-³¹P-NMR analyses. This method offers advantages such as simplicity workup with the green aspects by avoiding expensive catalysts and toxic solvents, good yields, short reaction times.

Keywords: amino acids esters, α-aminophosphonates, H₆P₂W₁₈O₆₂.14H₂O catalyst, green chemistry

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1013 Effect of Chemical Fertilizer on Plant Growth-Promoting Rhizobacteria in Wheat

Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline

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The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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1012 Influence of Bacterial Biofilm on the Corrosive Processes in Electronic Equipment

Authors: Iryna P. Dzieciuch, Michael D. Putman

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Humidity is known to degrade Navy ship electronic equipment, especially in hot moist environments. If left untreated, it can cause significant and permanent damage. Even rigorous inspection and frequent clean-up would not prevent further equipment contamination and degradation because of the constant presence of favorable growth conditions for many microorganisms. Generally, relative humidity levels of less than 60% will inhibit corrosion in electronic equipment, but because NAVY electronics often operate in hot and humid environments, prevention via dehumidification is not always possible. Currently, there is no defined research that fully describes key mechanisms which cause electronics and its coating degradation. The corrosive action of most bacteria is mainly developed through (i) mycelium adherence to the metal plates, (ii) facilitation the formation of pitting areas, (iii) production of organic acids such as citric, iso-citric, cis-aconitic, alpha-ketoglutaric, which are corrosive to electronic equipment and its components. Our approach studies corrosive action in electronic equipment: circuit-board, wires and connections that are exposed in the humid environment that gets worse during condensation. In our new approach the technical task is built on work with the bacterial communities in public areas, bacterial genetics, bioinformatics, biostatistics and Scanning Electron Microscopy (SEM) of corroded circuit boards. Based on these methods, we collect and examine environmental samples from biofilms of the corroded and non-corroded sites, where bacterial contamination of electronic equipment, such as machine racks and shore boats, is an ongoing concern. Sample collection and sample analysis is focused on addressing the key questions identified above through the following tasks: laboratory sample processing and evaluation under scanning electron microscopy, initial sequencing and data evaluation; bioinformatics and data analysis. Preliminary results from scanning electron microscopy (SEM) have revealed that metal particulates and alloys in corroded samples consists mostly of Tin ( < 40%), Silicon ( < 4%), Sulfur ( < 1%), Aluminum ( < 2%), Magnesium ( < 2%), Copper ( < 1%), Bromine ( < 2%), Barium ( <1%) and Iron ( < 2%) elements. We have also performed X 12000 magnification of the same sites and that proved existence of undisrupted biofilm organelles and crystal structures. Non-corrosion sites have revealed high presence of copper ( < 47%); other metals remain at the comparable level as on the samples with corrosion. We have performed X 1000 magnification on the non-corroded at the sites and have documented formation of copper crystals. The next step of this study, is to perform metagenomics sequencing at all sites and to compare bacterial composition present in the environment. While copper is nontoxic to the living organisms, the process of bacterial adhesion creates acidic environment by releasing citric, iso-citric, cis-aconitic, alpha-ketoglutaric acidics, which in turn release copper ions Cu++, which that are highly toxic to the bacteria and higher order living organisms. This phenomenon, might explain natural “antibiotic” properties that are lacking in elements such as tin. To prove or deny this hypothesis we will use next - generation sequencing (NGS) methods to investigate types and growth cycles of bacteria that from bacterial biofilm the on corrosive and non-corrosive samples.

Keywords: bacteria, biofilm, circuit board, copper, corrosion, electronic equipment, organic acids, tin

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1011 Advanced Biosensor Characterization of Phage-Mediated Lysis in Real-Time and under Native Conditions

Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka

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Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.

Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68

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1010 Analysis of Non-Coding Genome in Streptococcus pneumoniae for Molecular Epidemiology Typing

Authors: Martynova Alina, Lyubov Buzoleva

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Streptococcus pneumoniae is the causative agent of pneumonias and meningitids throught all the world. Having high genetic diversity, this microorganism can cause different clinical forms of pneumococcal infections and microbiologically it is really difficult diagnosed by routine methods. Also, epidemiological surveillance requires more developed methods of molecular typing because the recent method of serotyping doesn't allow to distinguish invasive and non-invasive isolates properly. Non-coding genome of bacteria seems to be the interesting source for seeking of highly distinguishable markers to discriminate the subspecies of such a variable bacteria as Streptococcus pneumoniae. Technically, we proposed scheme of discrimination of S.pneumoniae strains with amplification of non-coding region (SP_1932) with the following restriction with 2 types of enzymes of Alu1 and Mn1. Aim: This research aimed to compare different methods of typing and their application for molecular epidemiology purposes. Methods: we analyzed population of 100 strains of S.pneumoniae isolated from different patients by different molecular epidemiology methods such as pulse-field gel electophoresis (PFGE), restriction polymorphism analysis (RFLP) and multilolocus sequence typing (MLST), and all of them were compared with classic typing method as serotyping. The discriminative power was estimated with Simpson Index (SI). Results: We revealed that the most discriminative typing method is RFLP (SI=0,97, there were distinguished 42 genotypes).PFGE was slightly less discriminative (SI=0,95, we identified 35 genotypes). MLST is still the best reference method (SI=1.0). Classic method of serotyping showed quite weak discriminative power (SI=0,93, 24 genotypes). In addition, sensivity of RFLP was 100%, specificity was 97,09%. Conclusion: the most appropriate method for routine epidemiology surveillance is RFLP with non-coding region of Streptococcsu pneumoniae, then PFGE, though in some cases these results should be obligatory confirmed by MLST.

Keywords: molecular epidemiology typing, non-coding genome, Streptococcus pneumoniae, MLST

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1009 Effect of Treated Grey Water on Bacterial Concrete

Authors: Deepa T., Inchara S. R., Venkatesh S. V., Seema Tharannum

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Concrete is the most widely used structural material. It is usually made using locally available materials. However, concrete has low tensile strength and may crack in the early days with exothermic hydration, for which water is essential. To address the increased construction water demand, treated greywater may be used. Bacillus subtilis bacteria that form endospores is the biological agent considered in this study for biomineralization or Microbially Induced Calcite Precipitation (MICP) technique to heal cracks. Treated grey water which is obtained from STP of PES University, opted in place of Potable water, which had qualities within the standard range as per codal provisions. In this work, M30 grade conventional concrete is designed using OPC 53-grade cement, manufactured sand, natural coarse aggregates, and potable water. Conventional concrete (CC), bacterial concrete with potable water (BS), and treated grey water concrete (TGWBS) are the three different concrete specimens cast. Experimental studies such as the strength test and the surface hardness test are performed on conventional and bacterial concrete samples after 7, 28, and 56 days of curing. Concrete cubes are subjected to a temperature of 50° C to investigate the effect of higher temperature. Cracked cube specimens are observed for self-healing -as well as microstructure analysis with Scanning Electron Microscope (SEM), Energy Dispersive X-Ray Analysis (EDAX), and X-Ray Diffraction Analysis (XRD). Noticeable calcium salt deposition is observed on the surface of the BS and TGWBS cracked specimen. Surface hardness and the EDAX test gave promising results on the advantage of using spore-forming bacteria in concrete. This is followed by the strength gained in compression and flexure. Results also indicate that treated grey water can be a substitute for potable water in concrete.

Keywords: Bacillus subtilis concrete, microstructure, temperature, treated greywater

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1008 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

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1007 Selection of a Potential Starter Culture for Milk Fermentation

Authors: Stephen Olusanmi Akintayo, Ilesanmi Fadahunsi

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The ability of Lactic acid bacteria (LAB) to grow and survive in milk is being exploited in industrial and biotechnological applications. Although considerable studies have been reported on the fermentation of milk, however, not so much work has been documented on the selection of LAB strains from milk of the Nigerian local cattle breeds for their starter culture potentials. A total of 110 LAB were isolated from raw milk of Sokoto gudali cattle breed. The isolates were screened for their proteolytic activities on skimmed milk media with isolates A07, F06 and A01 showing the highest zone of clearance of 18.5mm, 18.5mm, and 18.0mm respectively and were selected for the studies of their growth in different constituents of milk. A01, F06, and A07 were identified as Pediococcus acidilactici, Lactococcus raffinolactis, and Leuconostoc mesenteriodes respectively using cultural, biochemical, physiological and molecular characterization techniques. Leuconostoc mesenteriodes showed the highest growth in all the milk components that were used in this study. The three LAB species selected showed a growth range of 6.46 log cfu/ml to 10.91 log cfu/ml in lactose with Leuconostoc mesenteriodes showing the highest growth of 10.91 log cfu/ml while Pediococcus acidilactici recorded the lowest growth of 9.78 log cfu/ml. In medium containing leucine as the only amino acid, the viable counts of Pediococcus acidilactici, Lactococcus raffinolactis and Leuconostoc mesenteriodes in log cfu/ml at zero hour were 6.39, 6.36 and 6.38 respectively which increased to 9.31 log cfu/ml, 9.21 log cfu/ml, 9.92 log cfu/ml respectively after 24 hours. Similarly, in all other substrates (casein, lysine, glutamic acid, aspartic acid, stearic acid and oleic acid ) tested in this study, Leuconostoc mesenteriodes showed the highest growth. It was observed that the highest quantity of lactic acid (15.31mg/ml) was produced by Leuconostoc mesenteriodes. The same trend was also observed in the production of diacetyl and hydrogen peroxide by the three tested microorganisms. Due to its ability to grow maximally in milk components, Leuconostoc mesenteriodes shows potential as starter culture for milk fermentation.

Keywords: Leuconostoc mesenteriodes, lactic acid bacteria, Sokoto gudali, starter culture

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1006 Analysis of Resistance and Virulence Genes of Gram-Positive Bacteria Detected in Calf Colostrums

Authors: C. Miranda, S. Cunha, R. Soares, M. Maia, G. Igrejas, F. Silva, P. Poeta

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The worldwide inappropriate use of antibiotics has increased the emergence of antimicrobial-resistant microorganisms isolated from animals, humans, food, and the environment. To combat this complex and multifaceted problem is essential to know the prevalence in livestock animals and possible ways of transmission among animals and between these and humans. Enterococci species, in particular E. faecalis and E. faecium, are the most common nosocomial bacteria, causing infections in animals and humans. Thus, the aim of this study was to characterize resistance and virulence factors genes among two enterococci species isolated from calf colostrums in Portuguese dairy farms. The 55 enterococci isolates (44 E. faecalis and 11 E. faecium) were tested for the presence of the resistance genes for the following antibiotics: erythromicyn (ermA, ermB, and ermC), tetracycline (tetL, tetM, tetK, and tetO), quinupristin/dalfopristin (vatD and vatE) and vancomycin (vanB). Of which, 25 isolates (15 E. faecalis and 10 E. faecium) were tested until now for 8 virulence factors genes (esp, ace, gelE, agg, cpd, cylA, cylB, and cylLL). The resistance and virulence genes were performed by PCR, using specific primers and conditions. Negative and positive controls were used in all PCR assays. All enterococci isolates showed resistance to erythromicyn and tetracycline through the presence of the genes: ermB (n=29, 53%), ermC (n=10, 18%), tetL (n=49, 89%), tetM (n=39, 71%) and tetK (n=33, 60%). Only two (4%) E. faecalis isolates showed the presence of tetO gene. No resistance genes for vancomycin were found. The virulence genes detected in both species were cpd (n=17, 68%), agg (n=16, 64%), ace (n=15, 60%), esp (n=13, 52%), gelE (n=13, 52%) and cylLL (n=8, 32%). In general, each isolate showed at least three virulence genes. In three E. faecalis isolates was not found virulence genes and only E. faecalis isolates showed virulence genes for cylA (n=4, 16%) and cylB (n=6, 24%). In conclusion, these colostrum samples that were consumed by calves demonstrated the presence of antibiotic-resistant enterococci harbored virulence genes. This genotypic characterization is crucial to control the antibiotic-resistant bacteria through the implementation of restricts measures safeguarding public health. Acknowledgements: This work was funded by the R&D Project CAREBIO2 (Comparative assessment of antimicrobial resistance in environmental biofilms through proteomics - towards innovative theragnostic biomarkers), with reference NORTE-01-0145-FEDER-030101 and PTDC/SAU-INF/30101/2017, financed by the European Regional Development Fund (ERDF) through the Northern Regional Operational Program (NORTE 2020) and the Foundation for Science and Technology (FCT). This work was supported by the Associate Laboratory for Green Chemistry - LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and UIDP/50006/2020).

Keywords: antimicrobial resistance, calf, colostrums, enterococci

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1005 Co-Immobilization of Palladium Nanoparticles and Polyoxometalate into the Cavities of the Mesocellular Foams: A Biomimetic Cooperative Catalytic System for Aerobic Oxidation of Alcohols under Green Conditions

Authors: Saeed Chehri, Sirvan Moradi, Amin Rostami

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Cooperative catalyst systems have been developed as highly promising sustainable alternatives to traditional catalysts. In these catalysts, two or more catalytic centers cooperate to reduce the energy of chemical transformations. In nature, such systems are abundantly seen in metalloenzymes that use metal and an organic cofactor. We have designed a reusable cooperative catalyst oxidation system consisting of palladium nanoparticles and polyoxometalate. This biomimetic cooperative catalytic system was synthesized by the stepwise immobilization of palladium nanoparticlesandpolyoxometalateinto the same cavity of siliceous mesocellularfoams (Pd-POM@MCF)and wascharacterizedby SEM, EDX, FT-IR, TGAand ICP techniques. POM-Pd@MCF/HQexhibits high activity toward aerobic oxidation of alcohols to the corresponding carbonyl compoundsin water solvent at room temperature. The major novelties and advantages of this oxidation method are as follows: (i) this is the first report of the co-immobilization of polyoxometalateand palladium for use as a robust and highlyefficient heterogeneouscooperative oxidative nanocatalyst system for aerobic oxidation of alcohols, (ii) oxidation of alcoholswere performed using an ideal oxidant with good to high yields in a green solvent at ambient temperature and (iii) the immobilization of the oxygen-activating catalyst(polyoxometalate) and oxidizing catalyst (Pd) onto MCF provide practical cooperative catalyst the system that can be reused several times without a significant loss of activity (vi) the methodsconform to several of the guiding principles of green chemistry.

Keywords: palladium nanoparticles, polyoxometalate, reusable cooperative catalytic system, biomimetic oxidation reaction

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1004 The Effect of Colloidal Metals Nanoparticles on Quarantine Bacterium - Clavibacter michiganensis Ssp. sepedonicus

Authors: Włodzimierz Przewodowski, Agnieszka Przewodowska

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Colloidal metal nanoparticles have drawn increasing attention in the field of phytopathology because of their unique properties and possibilities of applications. Their antibacterial activity, no induction of the development of pathogen resistance and the ability to penetrate most of biological barriers make them potentially useful in the fighting against dangerous pathogens. These properties are very important in the case of protection of strategic crops in the world, like potato - fourth crop in the world - which is host to numerous pathogenic microorganisms causing serious diseases, significantly affecting yield and causing the economic losses. One of the most important and difficult to reduce pathogen of potato plant is quarantine bacterium Clavibacter michiganensis ssp. sepedonicus (Cms) responsible for ring rot disease. Control and detection of these pathogens is very complicated. Application of healthy, certified seed material as well as hygiene in potato production and storage are the most efficient ways of preventing of ring rot disease. Currently used disinfectants and pesticides, have many disadvantages, such as toxicity, low efficiency, selectivity, corrosiveness, and the inability to eliminate the pathogens in potato tissue. In this situation, it becomes important to search for new formulations based on components harmful to health, yet efficient, stable during prolonged period of time and a with wide range of biocide activity. Such capabilities are offered by the latest generation of biocidal nanoparticles such as colloidal metals. Therefore the aim of the presented research was to develop newly antibacterial preparation based on colloidal metal nanoparticles and checking their influence on the Cms bacteria. Our preliminary results confirmed high efficacy of the nano-colloids in controlling the this selected pathogen.

Keywords: clavibacter michiganensis ssp. sepedonicus, colloidal metal nanoparticles, phytopathology, bacteria

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1003 Thermochemical Study of the Degradation of the Panels of Wings in a Space Shuttle by Utilization of HSC Chemistry Software and Its Database

Authors: Ahmed Ait Hou

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The wing leading edge and nose cone of the space shuttle are fabricated from a reinforced carbon/carbon material. This material attains its durability from a diffusion coating of silicon carbide (SiC) and a glass sealant. During re-entry into the atmosphere, this material is subject to an oxidizing high-temperature environment. The use of thermochemical calculations resulting at the HSC CHEMISTRY software and its database allows us to interpret the phenomena of oxidation and chloridation observed on the wing leading edge and nose cone of the space shuttle during its mission in space. First study is the monitoring of the oxidation reaction of SiC. It has been demonstrated that thermal oxidation of the SiC gives the two compounds SiO₂(s) and CO(g). In the extreme conditions of very low oxygen partial pressures and high temperatures, there is a reaction between SiC and SiO₂, leading to SiO(g) and CO(g). We had represented the phase stability diagram of Si-C-O system calculated by the use of the HSC Chemistry at 1300°C. The principal characteristic of this diagram of predominance is the line of SiC + SiO₂ coexistence. Second study is the monitoring of the chloridation reaction of SiC. The other problem encountered in addition to oxidation is the phenomenon of chloridation due to the presence of NaCl. Indeed, after many missions, the leading edge wing surfaces have exhibited small pinholes. We have used the HSC Chemistry database to analyze these various reactions. Our calculations concorde with the phenomena we announced in research work resulting in NASA LEWIS Research center.

Keywords: thermochchemicals calculations, HSC software, oxidation and chloridation, wings in space

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1002 Characterization of Emissions from the open burning of Municipal Solid Waste (MSW) under Tropical Environment

Authors: Anju Elizbath Peter, S. M. Shiva Nagendra, Indumathi M.Nambi

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The deliberate fires initiated by dump managers and human scavengers to reduce the volume of waste and recovery of valuable metals/materials are common at municipal solid waste (MSW) disposal sites in developed country. A large amount of toxic gases released due to this act is responsible for the deterioration of regional and local air quality, which causes visibility impairment and acute respiratory diseases. The present study was aimed at the characterization of MSW and emission characteristics of burning of MSW in the laboratory. MSW samples were collected directly from the one of the open dumpsite located in Chennai city. Solid waste sampling and laboratory analysis were carried out according to American Society of Testing and Materials (ASTM) standards. Results indicated the values of moisture content, volatile solids (VS) and calorific values of solid waste samples were 16.67%,8%,9.17MJ/kg, respectively. The elemental composition showed that the municipal solid waste contains 25.84% of carbon, 3.69% of hydrogen, 1.57% of nitrogen and 0.26% of sulphur. The calorific value of MSW was found to be 9.17 MJ/Kg which is sufficient to facilitate self-combustion of waste. The characterization of emissions from the burning of 1 kg of MSW in the test chamber showed a total of 90 mg/kg of PM10 and 243 mg/kg of PM2.5. The current research study results will be useful for municipal authorities to formulate guideline and policy structure regarding the MSW management to reduce the impact of air emissions at an open dump site.

Keywords: characterization, MSW, open burning, PM10, PM2.5

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