Search results for: bacterial biomass

Authors: Blessing Atim Aderibigbe

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A series of amine/ester-linked hybrid compounds containing pharmacophores, such as ursolic acid, oleanolic acid, ferrocene and bisphosphonates, were synthesized in an attempt to develop potent antibacterial and anticancer agents. Their structures were analyzed and confirmed using Nuclear Magnetic Resonance, Fourier Transform Infrared Spectroscopy, and mass spectroscopy. All the synthesized hybrid compounds were evaluated for their antibacterial activities against eleven selected bacterial strains using a serial dilution method. Some of the compounds displayed significant antibacterial activity against most of the bacterial and fungal strains. In addition, the in vitro cytotoxicity of these compounds was also performed against selected cancer cell lines. Some of the compounds were also found to be more active than their parent compounds, revealing the efficacy of designing hybrid molecules using plant-based bioactive agents.

Keywords: ursolic acid, hybrid drugs, oleanolic acid, bisphosphonates

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Authors: Keigo Ide, Toru Maruyama, Michihiro Ito, Hiroyuki Fujimura, Yoshikatu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

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Understanding environmental system is one of the important tasks. In recent years, conservation of coral environments has been focused for biodiversity issues. The damage of coral reef under environmental impacts has been observed worldwide. However, the casual relationship between damage of coral and environmental impacts has not been clearly understood. On the other hand, structure/diversity of marine bacterial community may be relatively robust under the certain strength of environmental impact. To evaluate the coral environment conditions, it is necessary to investigate relationship between marine bacterial composition in coral reef and environmental factors. In this study, the Time Scale Network Analysis was developed and applied to analyze the marine environmental data for investigating the relationship among coral, bacterial community compositions and environmental factors. Seawater samples were collected fifteen times from November 2014 to May 2016 at two locations, Ishikawabaru and South of Sesoko in Sesoko Island, Okinawa. The physicochemical factors such as temperature, photosynthetic active radiation, dissolved oxygen, turbidity, pH, salinity, chlorophyll, dissolved organic matter and depth were measured at the coral reef area. Metagenome and metatranscriptome in seawater of coral reef were analyzed as the biological factors. Metagenome data was used to clarify marine bacterial community composition. In addition, functional gene composition was estimated from metatranscriptome. For speculating the relationships between physicochemical and biological factors, cross-correlation analysis was applied to time scale data. Even though cross-correlation coefficients usually include the time precedence information, it also included indirect interactions between the variables. To elucidate the direct regulations between both factors, partial correlation coefficients were combined with cross correlation. This analysis was performed against all parameters such as the bacterial composition, the functional gene composition and the physicochemical factors. As the results, time scale network analysis revealed the direct regulation of seawater temperature by photosynthetic active radiation. In addition, concentration of dissolved oxygen regulated the value of chlorophyll. Some reasonable regulatory relationships between environmental factors indicate some part of mechanisms in coral reef area.

Keywords: coral environment, marine microbiology, network analysis, omics data analysis

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Authors: Andres Diaz Garcia, Ana Maria Ceballos Rojas, Duvan Albeiro Millan Montano

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This paper describes the initial technological development stages in the area of liquid fermentation required to reach the quantities of biomass of the biofertilizer microorganism Rhizobium sp. strain B02, for the application of the unitary stages downstream at laboratory scale. In the first stage, the adjustment and standardization of the fermentation process in conventional batch mode were carried out. In the second stage, various fed-batch and continuous fermentation strategies were evaluated in 10L-bioreactor in order to optimize the yields in concentration (Colony Forming Units/ml•h) and biomass (g/l•h), to make feasible the application of unit operations downstream of process. The growth kinetics, the evolution of dissolved oxygen and the pH profile generated in each of the strategies were monitored and used to make sequential adjustments. Once the fermentation was finished, the final concentration and viability of the obtained biomass were determined and performance parameters were calculated with the purpose of select the optimal operating conditions that significantly improved the baseline results. Under the conditions adjusted and standardized in batch mode, concentrations of 6.67E9 CFU/ml were reached after 27 hours of fermentation and a subsequent noticeable decrease was observed associated with a basification of the culture medium. By applying fed-batch and continuous strategies, significant increases in yields were achieved, but with similar concentration levels, which involved the design of several production scenarios based on the availability of equipment usage time and volume of required batch.

Keywords: biofertilizer, liquid fermentation, Rhizobium sp., standardization of processes

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Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis

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The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.

Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis

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Authors: Jerry John T. M., Sylas V. P., Shijo Joy

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Hydrogen is the most essential gas that can be used for many purposes. During the production of hydrogen using raw materials like Soil and leftover cooked rice water (kanjivellam), the major by-product formed is water. Soil is collected from three different places in kottayam district: Kallara, Meenachilar, and Athirampuzha. Collected samples are mixed with rice water and tested for traces of hydrogen using a biohydrogen sensor after 72 hours. The result was the presence of hydrogen in all the 3 samples. After streaking, PCR and gel electrophoresis detected the bacteria which produced the hydrogen. RGCB Thiruvananthapuram conducted the sequencing of the PCR resultant. And identified the bacterial strains. Five variants of Bacillus bacteria ( (1) Bacillus cereus strain JTM GenBank: OP278839.1 (2) Bacillus toyonensis strain JTM2 GenBank: OP278841.1 (3) Bacillus anthracis strain JTM_SR2989-3-R_H08 GenBank: OP278960.1 (4) Bacillus thuringiensis strain JRY1 GenBank: OP278976.1 (5) Bacillus anthracis strain JTM_SR2989-3-F_H07 GenBank: OP278959.1 ) are identified and successfully registered in NCBI Gen bank. These Bacillus bacteria are major types of Rhizobacteria that can form spores and can survive in the soil for a long time period under harsh environmental conditions. Also, plant growth is enhanced by PGPR (Plant growth promoting rhizobacteria) through the induction of systemic resistance, antibiosis, and competitive omission. The molecular sequencing was submitted to the NCBI Gen Bank, and the accession numbers were allotted for the bacterial cultures.

Keywords: bio hydrogen production, bacterial bio hydrogen production, plant related to bacillus bacteria., bacillus bacteria study

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Authors: Shayma Munqith Al-Baker, Fadhl Ahmed Saeed Al-Gasha’a, Samira Hamid Hanash, Ahmed Ali Al-Hazmi

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A total of 200 samples (180 fecal materials and 20 organ samples) were collected from (5 different poultry farms, 10 local poultry shops, 5 houses poultry, 5 Eggs stores shops and 5 hand slaughters centers) in Ibb city, Yemen, 2014. According to morphological, cultural, as well as biochemical characterization and serological tests, 59 29.5% isolates were identified as Salmonella spp. and all Salmonella isolates were categorized by serotype, which comprised of, 37 62.71% Salmonella Typhimurium serovar, 21 35.59%. Salmonella Enteritidis serovar and 11.69% Salmonella Heidelberg serovar. Antibiotic sensitivity test was done for bacterial isolates and the results showed there were clear differences in antibiotic resistant. Antimicrobial susceptibility of the isolates varies as follows: Ofloxacin 79.66%, Ciprofloxacin 67.80%, Colistin 59.32% and Gentamycin 52.54%. All of isolates were resistant to Erythromycin, Penicillin and Lincomycin. Antibacterial activity was done for both aqueous and ethanol extracts of Dodonaea viscosa plant by using well and disc diffusion assay. The results indicated that well diffusion assay had best results than disc diffusion assay, the highest inhibition zone was 22 mm for well diffusion and 15 mm for disc diffusion assay, the results observed that ethanol extract had best antibacterial effect than aqueous extract which the percentage of bacterial isolates affected with ethanol extract was 71.19% comparing with aqueous extract 28.81% by using disc diffusion assay, while the percentage of bacterial isolates affected with ethanol extract was 88.13% comparing with aqueous extract 52.54% by using will diffusion assay.

Keywords: Salmonella spp, Dodonaea viscosa, antimicrobial and salmonellosis

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Authors: Weicheng Luo, Wenzhi Zhao

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Plants growth in desert often suffered from stresses like water deficit, wind erosion and sand burial. Thus, plants in desert always have unique strategies to adapt these stresses. However, data regarding how clonal shrubs withstand wind erosion and sand burial in natural habitats remain relatively scarce. Therefore, we selected a common clonal shrub Calligonum arborescens to study the adaptive strategies of clonal plants to sand dune environment in a transitional zone of desert and Hexi Oasis of China. Our results show that sand burial is one of the essential prerequisites for the survival of C. arborescens rhizome fragments. Both the time and degrees of sand burial and wind erosion had significantly effects on clonal reproduction and growth of C. arborescens. With increasing burial depth, the number of ramets and biomass production significantly decreased. There is same change trend in severe erosion treatments. However, the number of ramets and biomass production significantly increased in moderate erosion treatments. Rhizome severed greatly decreased ramet number and biomass production under both sand burial and severe erosion treatments. That indicated that both sand burial and severe erosion had negative effects on the clonal growth of C. arborescens, but moderate wind erosion had positive effects. And rhizome connections alleviated the negative effects of sand burial and of severe erosion on the growth and performance of C. arborescens. Most fragments of C. arborescens grew in the directions of northeastern and southwestern. Ramet number and biomass, rhizome length and biomass in these two directions were significantly higher than those found in other directions. Interestingly, these directions were perpendicular to the prevailing wind direction. Distribution of C. arborescens differed in different habitats. The total number of individuals was significantly higher in inter-dune areas and on windward slopes than on the top and leeward slopes of dunes; more clonal ramets were produced on the top of dunes than elsewhere, and a few were found on leeward slopes. The mainly reason is that ramets on windward and top of dunes can easily suffered with moderated wind erosion which promoted clonal growth and reproduction of C. arborescens. These results indicated that C. arborescens adapted sand dune environment through directional growth and patchy distribution, and sand-burial and wind erosion were the key factors which led to the directional growth and patchiness of C. arborescens.

Keywords: adaptive strategy, Calligonum arborescens Litv, clonal fragment, desert-oasis transitional zone, sand burial and wind erosion

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Authors: Won Yeol Choi, Sangmo Kang

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The propulsion of a bacterial flagellum in a viscous fluid has attracted many interests in the field of biological hydrodynamics, but remains yet fully understood and thus still a challenging problem. In this study, therefore, we have numerically investigated the flow around a steadily rotating micro-sized spring to further understand such bacterial flagellum propulsion. Note that a bacterium gains thrust (propulsive force) by rotating the flagellum connected to the body through a bio motor to move forward. For the investigation, we convert the spring model from the micro scale to the macro scale using a similitude law (scale law) and perform simulations on the converted macro-scale model using a commercial software package, CFX v13 (ANSYS). To scrutinize the propulsion characteristics of the flagellum through the simulations, we make parameter studies by changing some flow parameters, such as the pitch, helical radius and rotational speed of the spring and the Reynolds number (or fluid viscosity), expected to affect the thrust force experienced by the rotating spring. Results show that the propulsion characteristics depend strongly on the parameters mentioned above. It is observed that the forward thrust increases in a linear fashion with either of the rotational speed or the fluid viscosity. In addition, the thrust is directly proportional to square of the helical radius and but the thrust force is increased and then decreased based on the peak value to the pitch. Finally, we also present the appropriate flow and pressure fields visualized to support the observations.

Keywords: fluid viscosity, hydrodynamics, similitude, propulsive force

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Authors: Asma Aid, Samira Amokrane, Djamel Nibou, Hadj Mekatel

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The marine Enteromorpha Compressa (L.) (ECL) biomass was used as a low-cost biological adsorbent for the removal of Cr⁶⁺, Co²⁺ and Ni²⁺ ions from artificially contaminated aqueous solutions. The operating variables pH, the initial concentration C₀, the solid/liquid ratio R and the temperature T were studied. A full factorial experimental design technique enabled us to obtain a mathematical model describing the adsorption of Cr⁶⁺, Co²⁺ and Ni²⁺ ions and to study the main effects and interactions among operational parameters. The equilibrium isotherm has been analyzed by Langmuir, Freundlich, and Dubinin-Radushkevich models; it has been found that the adsorption process follows the Langmuir model for the used ions. Kinetic studies showed that the pseudo-second-order model correlates our experimental data. Thermodynamic parameters showed the endothermic heat of adsorption and the spontaneity of the adsorption process for Cr⁶⁺ ions and exothermic heat of adsorption for Co²⁺ and Ni²⁺ ions.

Keywords: enteromorpha Compressa, adsorption process, Cr⁶⁺, Co²⁺ and Ni²⁺, equilibrium isotherm

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Authors: Azna Zuberi

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Biofilm is a sessile bacterial accretion in which bacteria adapts different physiological and morphological behavior from planktonic form. It is the root cause of about 80% microbial infections in human. Among them, E. coli biofilms are most prevalent in medical devices associated nosocomial infections. The objective of this study was to inhibit biofilm formation by targeting LuxS gene, involved in quorum sensing using CRISPRi. luxS is a synthase, involved in the synthesis of Autoinducer-2(AI-2), which in turn guides the initial stage of biofilm formation. To implement CRISPRi system, we have synthesized complementary sgRNA to target gene sequence and co-expressed with dCas9. Suppression of luxS was confirmed through qRT-PCR. The effect of luxS gene on biofilm inhibition was studied through crystal violet assay, XTT reduction assay and scanning electron microscopy. We conclude that CRISPRi system could be a potential strategy to inhibit bacterial biofilm through mechanism base approach.

Keywords: biofilm, CRISPRi, luxS, microbial

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Authors: Thais O. de Paula, Marjorie R. A. Sarmiento, Francis M. Borges, Alessandra B. Ferreira-Machado, Juliana A. Resende, Dioneia E. Cesar, Vania L. Silva, Claudio G. Diniz

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Obesity is currently a worldwide public health threat, being considered a pandemic multifactorial disease related to the human gut microbiota (GM). Add to that GM is considered an important reservoir of antimicrobial resistance genes (ARG) and little is known on GM and ARG in obesity, considering the altered physiology and xenobiotics intake. As regional and social behavior may play important roles in GM modulation, and most of the studies are based on small sample size and various methodological approaches resulting in difficulties for data comparisons, this study was focused on the investigation of GM bacterial diversity in obese (OB), overweight (OW) and eutrophic individuals (ET) considering their nutritional, clinical and social characteristics; and comparative screening of AGR related to their physiology and xenobiotics intake. Microbial community was accessed by FISH considering phyla as a taxonomic level, and PCR-DGGE followed by dendrograms evaluation (UPGMA method) from fecal metagenome of 72 volunteers classified according to their body mass index (BMI). Nutritional, clinical, social parameters and xenobiotics intake were recorded for correlation analysis. The fecal metagenome was also used as template for PCR targeting 59 different ARG. Overall, 62% of OB were hypertensive, and 12% or 4% were, regarding the OW and ET individuals. Most of the OB were rated as low income (80%). Lower relative bacterial densities were observed in the OB compared to ET for almost all studied taxa (p < 0.05) with Firmicutes/Bacteroidetes ratio increased in the OB group. OW individuals showed a bacterial density representative of GM more likely to the OB. All the participants were clustered in 3 different groups based on the PCR-DGGE fingerprint patterns (C1, C2, C3), being OB mostly grouped in C1 (83.3%) and ET mostly grouped in C3 (50%). The cluster C2 showed to be transitional. Among 27 ARG detected, a cluster of 17 was observed in all groups suggesting a common core. In general, ARG were observed mostly within OB individuals followed by OW and ET. The ratio between ARG and bacterial groups may suggest that AGR were more related to enterobacteria. Positive correlations were observed between ARG and BMI, calories and xenobiotics intake (especially use of sweeteners). As with nutritional and clinical characteristics, our data may suggest that GM of OW individuals behave in a heterogeneous pattern, occasionally more likely to the OB or to the ET. Regardless the regional and social behaviors of our population, the methodological approaches in this study were complementary and confirmatory. The imbalance of GM over the health-disease interface in obesity is a matter of fact, but its influence in host's physiology is still to be clearly elucidated to help understanding the multifactorial etiology of obesity. Although the results are in agreement with observations that GM is altered in obesity, the altered physiology in OB individuals seems to be also associated to the increased xenobiotics intake and may interfere with GM towards antimicrobial resistance, as observed by the fecal metagenome and ARG screening. Support: FAPEMIG, CNPQ, CAPES, PPGCBIO/UFJF.

Keywords: antimicrobial resistance, bacterial diversity, gut microbiota, obesity

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Authors: Thirunavoukkarasu Manikkannan, Karpanai Selvan Balasubramanian

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Textile industry is the one of the most important industrial sector in India. It accounts for 5% of total Gross Domestic Product (GDP) in the country. A Textile industry consumes large quantities of water (~250 m3/ton of product) and they generate almost ~90% of wastewater from its consumption. The problem is alarming and requires proper treatment process to acquire dual benefit of Zero Liquid Discharge and no contamination to the environment. Here we describe the process by which the textile wastewater can be reused. We have collected the textile wastewater in and around Ayyampettai area of Tamilnadu, India. Among different microalgal strains used, Desodesmus sp. collected at Manali, Chennai, Tamilnadu, India was able to lessen the colour of the waste water in 12-15 hrs of its growth, COD around 81.7%, Dissolved solid reduction was 28 ± 0.5 %, Suspended solid was reduced to 40.5 ± 0.3 %, Dye degradation was 50-78%. Further, Desodesmus sp. able to achieve the biomass of 0.9 ± 0.2 g/L (dry weight) in two weeks’ time, the Chl a content was 11 mg/L. It infers that this algal strain able to utilize the textile wastewater as source for growth and algal biomass production.

Keywords: Desodesmus sp., microalgae, textile, treatment, wastewater

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Authors: Ali Fadhel Ahmed, Tuqa Abdulkareem Hameed

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Moringa oleifera (leaves and seeds) ethanolic and aqueous extracts were tested for antibacterial activity. The effect of plant extracts on three types of bacterial species: Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, was investigated. Using the agar well diffusion method, ethanolic extracts of Moringa oleifera demonstrated a significant antibacterial effect on the forty tested bacterial strains. Seed-induced inhibition zones (ethanolic extracts)were ranged from16 to 24 mm in diameter against S. aureus, respectively, whileE. coli and K. pneumonia had no effect. Gram-positive and Gram-negative bacteria were not affected by alcoholic and aqueous plant leaf extracts. The purpose of this present study was to look at the cytotoxic effects of M.Oleifera plant (alcoholic extracts).

Keywords: moringa oleifera, escherichia coli, klebsiella pneumoniae, staphylococcus aureus

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Authors: Ayesha Irum, Sadia Mumtaz, Abdul Rehman, Iffat Naz, Safia Ahmed

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The present study was conducted to evaluate the potential applicability of biological trickling filter system for the treatment of simulated textile wastewater containing reactive azo dyes with bacterial consortium under non-sterile conditions. The percentage decolorization for the treatment of wastewater containing structurally different dyes was found to be higher than 95% in all trials. The stable bacterial count of the biofilm on stone media of the trickling filter during the treatment confirmed the presence, proliferation, dominance and involvement of the added microbial consortium in the treatment of textile wastewater. Results of physicochemical parameters revealed the reduction in chemical oxygen demand (58.5-75.1%), sulphates (18.9-36.5%), and phosphates (63.6-73.0%). UV-Visible and FTIR spectroscopy confirmed decolorization of dye containing wastewater was the ultimate consequence of biodegradation. Toxicological studies revealed the nontoxic nature of degradative metabolites.

Keywords: biodegradation, textile dyes, waste water, trickling filters

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Authors: Ismail S. Bostanci, Ebru Akkaya

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Microalgae-derived biodiesel, fertilizer or industrial chemicals' production with wastewater has great potential. Especially water from a municipal wastewater treatment plant is a very important nutrient source for biofuel production. Microalgae biomass production in open ponds system is lower cost culture systems. There are many hurdles for commercial algal biomass production in large scale. One of the important technical bottlenecks for microalgae production in open system is culture contamination. The algae culture contaminants can generally be described as invading organisms which could cause pond crash. These invading organisms can be competitors, parasites, and predators. Contamination is unavoidable in open systems. Potential contaminant organisms are already inoculated if wastewater is utilized for algal biomass cultivation. Especially, it is important to control contaminants to retain in acceptable level in order to reach true potential of algal biofuel production. There are several contamination management methods in algae industry, ranging from mechanical, chemical, biological and growth condition change applications. However, none of them are accepted as a suitable contamination control method. This experiment describes an innovative contamination control method, 'Recirculated Sedimentation Method', to manage contamination to avoid pond cash. The method can be used for the production of algal biofuel, fertilizer etc. and algal wastewater treatment. To evaluate the performance of the method on algal culture, an experiment was conducted for 90 days at a lab-scale raceway (60 L) reactor with the use of non-sterilized and non-filtered wastewater (secondary effluent and centrate of anaerobic digestion). The application of the method provided the following; removing contaminants (predators and diatoms) and other debris from reactor without discharging the culture (with microscopic evidence), increasing raceway tank’s suspended solids holding capacity (770 mg L-1), increasing ammonium removal rate (29.83 mg L-1 d-1), decreasing algal and microbial biofilm formation on inner walls of reactor, washing out generated nitrifier from reactor to prevent ammonium consumption.

Keywords: contamination control, microalgae culture contamination, pond crash, predator control

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Authors: Nebiyou Masebo

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The rapid conversion of an old aged agroforestry (AF) based agricultural system to monocropping farming system in southern Ethiopia is increasing. The consequence of this, combined with climate change, has been impaired biodiversity, soil microbial biomass carbon (MBC), and soil organic carbon (SOC). The AF system could curb such problems due it is an ecologically and economically sustainable strategies. This study was aimed to investigate different agroforestry practices (AFPs) on MBC and SOC in southern Ethiopia. Soil samples were collected from homegarden based agroforestry practice (HAFP), crop land based agroforestry practice (ClAFP), woodlot based agroforestry practice (WlAFP), and trees on soil and water conservation based agroforestry practice (TSWAFP) using two depth layer (0-30 & 30-60 cm) by systematic sampling. Moreover, woody species inventorywas also collected. The chloroform fumigation extraction method was employed to determine MBC from different AFP types. In this study, the value of MBC and SOC decreased significantly with soil depth (p< 0.05). Besides, AFP type, soil depth, woody species diversity, and key soil properties also strongly influenced MBC and SOC (p< 0.05). In this study, the MBC was the highest (786 mg kg⁻¹ soil) in HAFP, followed by WlAFP (592 mg kg⁻¹ soil), TSWAFP (421 mg kg⁻¹ soil), and ClAFP (357 mg kg⁻¹ soil). The highest mean value of SOC (43.5Mg C ha⁻¹) was recorded in HAFP, followed by WlAFP (35.1Mg C ha⁻¹), TSWAFP (22.3 Mg C ha⁻¹), while the lowest (21.8 Mg C ha⁻¹) was recorded in ClAFP. The HAFP had high woody species diversity, and the lowest was recorded in ClAFP. The finding indicated that SOC and MBC were significantly affected by land management practices, and HAFP has the potential to improve MBC and SOC through good management practices of AFP.

Keywords: agroforestry practices, microbial biomass carbon, soil carbon, rapid conversion

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Authors: Faycal Boughalleb, Raoudha Abdellaoui

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The effect of NaCl stress on root and leaf anatomy was investigated in three Astragalus species grown in 0-300 mM NaCl for 30 days under greenhouse conditions. Root cross section and cortex thickness was reduced under salt stress in both species while A. tenuifolius showed thinner cortex and the root cross section was unchanged. The epidermis stele thickness was unaffected by salinity in A. armatus and A. tenuifolius and was reduced in A. mareoticus with smaller xylem vessel size. In addition, vessel density and wall thickness of xylem was increased under salt conditions in the studies species. The entire lamina and mesophyll of the three species were thinner in salt-stressed plants. A. armatus and A. tenuifolius showed the higher thickness with increased size of the lower epidermis. NaCl (300 mM) reduced leaf water content by 41.5 % in A. mareoticus while it was unchanged in the other species. The size of the vascular bundle increased under salinity in A. tenuifolius leaves and it was unchanged in the other ones. A longer distance between leaf vascular bundle was occurred in A. mareoticus. The effects of NaCl on root and leaf ultrastructure are discussed in relation to the degree of salt resistance of these species. The unchanged biomass production under salt stress confirmed the higher tolerance oft A. tenuifolius to salinity. A. armatus was moderately salt tolerant with decrease of biomass production by 14.2 % while A. mareoticus was considered as salt sensitive plant when the decrease in biomass production reached 56.8%.

Keywords: Astragalus species, leaf ultrastructure, root anatomy, salt stress

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Authors: Ghazi Faisal Najmuldeen, Noridah Abdullah, Mimi Sakinah

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Bioethanol is an alcohol made by fermentation, mostly from carbohydrates, Cellulosic biomass, derived from non-food sources, such as castor shell waste, is also being developed as a feedstock for ethanol production Cellulose extracted from biomass sources is considered the future feedstock for many products due to the availability and eco-friendly nature of cellulose. In this study, castor shell (CS) biowaste resulted from the extraction of Castor oil from castor seeds was evaluated as a potential source of cellulose. The cellulose was extracted after pretreatment process was done on the CS. The pretreatment process began with the removal of other extractives from CS, then an alkaline treatment, bleaching process with hydrogen peroxide, and followed by a mixture of acetic and nitric acids. CS cellulose was analysed by infrared absorption spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). The result showed that the overall process was adequate to produce cellulose with high purity and crystallinity from CS waste. The cellulose was then hydrolyzed to produce glucose and then fermented to bioethanol.

Keywords: bioethanol, castor shell, cellulose, biowaste

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Authors: Debajit Kalita, Macmillan Nongkhlaw, S. R. Joshi

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Mercury (Hg) is a highly toxic heavy metal released both from naturally occurring volcanoes and anthropogenic activities like alkali and mining industries as well as biomedical wastes. Exposure to mercury is known to affect the nervous, gastrointestinal and renal systems. In the present study, a bacterial isolate identified using 16S rRNA marker as Paenibacillus jamilae PKR1 isolated from India’s largest sandstone-type uranium deposits, containing an average of 0.1% U3O8, was found to be resistance to Hg contamination under culture conditions. It showed strong hydrophobicity as revealed by SAT, MATH, PAT, SAA adherence assays. The Fourier Transform Infrared (FTIR) spectra showed the presence of hydroxyl, amino and carboxylic functional groups on the cell surface EPS which are known to contribute in the binding of metals. It is proposed that the characterized isolate tolerating up to 4.0mM of mercury provides scope for its application in bioremediation of mercury from contaminated sites.

Keywords: mercury, Domiasiat, uranium, paenibacillus jamilae, hydrophobicity, FTIR

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Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

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For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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Authors: Paweł Stączek, Tomasz Plech, Aleksandra Strzelczyk, Katarzyna Dzitko, Monika Wujec, Edyta Kuśmierz, Piotr Paneth, Agata Paneth

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In this contribution, we will describe the inhibitory potency of nine thiosemicarbazide derivatives against bacterial type IIA topoisomerases, their antibacterial profile, and molecular modeling evaluation. We have found that one of the tested compounds, 4-benzoyl-1-(2-methyl-furan-3-ylcarbonyl) thiosemicarbazide, remarkably inhibits the activity of S. aureus DNA gyrase with the IC50 below 5 μM. Besides, this compound displays antibacterial activity on Staphylococcus spp. and E. faecalis at non-cytotoxic concentrations in mammalian cells, with minimal inhibitory concentrations (MICs) values at 25 μg/mL. Based on the enzymatic and molecular modeling studies we propose two factors, i.e. geometry of molecule and hydrophobic/hydrophilic balance as important molecular properties for developing thiosemicarbazide derivatives as potent Staphylococcus aureus DNA gyrase inhibitors.

Keywords: bioactivity, drug design, topoisomerase, molecular modeling

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Authors: Manal Suleiman, George Abu-Aqil, Uraib Sharaha, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman, Mahmoud Huleihel

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Klebsiella pneumoniae is one of the most aggressive multidrug-resistant bacteria associated with human infections resulting in high mortality and morbidity. Thus, for an effective treatment, it is important to diagnose both the species of infecting bacteria and their susceptibility to antibiotics. Current used methods for diagnosing the bacterial susceptibility to antibiotics are time-consuming (about 24h following the first culture). Thus, there is a clear need for rapid methods to determine the bacterial susceptibility to antibiotics. Infrared spectroscopy is a well-known method that is known as sensitive and simple which is able to detect minor biomolecular changes in biological samples associated with developing abnormalities. The main goal of this study is to evaluate the potential of infrared spectroscopy in tandem with Random Forest and XGBoost machine learning algorithms to diagnose the susceptibility of Klebsiella pneumoniae to antibiotics within approximately 20 minutes following the first culture. In this study, 1190 Klebsiella pneumoniae isolates were obtained from different patients with urinary tract infections. The isolates were measured by the infrared spectrometer, and the spectra were analyzed by machine learning algorithms Random Forest and XGBoost to determine their susceptibility regarding nine specific antibiotics. Our results confirm that it was possible to classify the isolates into sensitive and resistant to specific antibiotics with a success rate range of 80%-85% for the different tested antibiotics. These results prove the promising potential of infrared spectroscopy as a powerful diagnostic method for determining the Klebsiella pneumoniae susceptibility to antibiotics.

Keywords: urinary tract infection (UTI), Klebsiella pneumoniae, bacterial susceptibility, infrared spectroscopy, machine learning

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Authors: Fitri Ayu, Nadia, Tanti, Putri, Fatkhan, Pasid Harlisa, Suparmi

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Acne is a skin abnormal conditions experienced by many teens, this is caused by various factors such as the climate is hot, humid and excessive sun exposure can aggravate acne because it will lead to excess oil production. Flavonoids form complex compounds against extracellular proteins that disrupt the integrity of bacterial cell membrane in a way denature bacterial cell proteins and bacterial cell membrane damage. This study aimed to test the antibacterial activity of corn silk extract with a concentration of 10 %, 20 %, 30 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 % and 100 % in vitro by measuring the inhibition of the growth of bacteria Propionibacterium acne, Staphylococcus aureus and Staphylococcus epidermis then compared with the standard antibiotic clindamycin. Extracts tested by Disk Diffusion Method, in which the blank disc soaked with their respective corn silk extract concentration for 15-30 minutes and then the medium of bacteria that have been planted with Propionibacterium acne, Staphylococcus aureus and Staphylococcus epidermis in the given disk that already contains extracts with various concentration. Incubated for 24 hours and then measured the growth inhibition zone Propionibacterium acne, Staphylococcus aureus and Staphylococcus epidermidis. Corn silk contains flavonoids, is shown by the test of flavonoids in corn silk extract by using a tube heating and without heating. Flavonoid in corn silk potentially as anti acne by inhibiting the growth of bacteria that cause acne. Corn silk extract concentration which has the highest antibacterial activity is then performed in a cream formulation and evaluation test of physical and chemical properties of the resulting cream preparation.

Keywords: antibacterial, flavonoid, corn silk, acne

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Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

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For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

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This publication presents a device for depolymerisation of plant substrates applicable to agricultural biogas plants and closed-chamber sewage treatment plants where sludge fermentation is bolstered with plant mass. The device consists of a tank with a cover equipped with a heating system, an inlet for the substrate, and an outlet for the depolymerised substrate. Within the tank, a magnet shaft encased in a spiral casing is attached, equipped on its upper end with an internal magnetic disc. A motoreducer is mounted on an external magnetic disc located on the centre of the cover. Depolymerisation of the plant substrate allows for substrate destruction at much lower power levels than by conventional means. The temperature within the reactor can be lowered by 40% in comparison to existing designs. During the depolymerisation process, free radicals are generated within the magnetic field, oxidizing the conditioned substrate and promoting biodegradation. Thus, the fermentation time in the fermenters is reduced by approximately 20%.

Keywords: depolymerisation, pre-treatment, biomass, fermentation

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Authors: Padmanabhan D, Kavitha S

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The purpose of the study is to investigate arsenic resistance ability of indigenous microflora and its ability to utilize arsenic species form containing water source. PS-6 potential arsenic tolerance bacterium was screened from thirty isolates from Pichavaram Mangroves of India having tolerance to grow up to 1000 mg/l of As (V) and 800 mg/l of As (III) and arsenic utilization ability of 98 % of As (V) and 97% of As (III) with initial concentration of 3-5 mg/l within 48 hrs. Optimum pH and temperature was found to be ~7-7.4 and 37°C. Active growth of PS-6 in minimal salt media (MSB) helps in cost effective biomass production. Dry weight analysis of PS-6 has shown significant difference in biomass when exposed to As (III) and As (V). Protein level study of PS-6 after exposing to As (V) and As (III) shown modification in total protein concentration and variation in SDS-PAGE pattern. PS-6 was identified as Bacillus licheniformis based on partially sequenced of 16S rRNA using NCBI Blast. Further investigation will help in using this potential bacterium as a well-grounded source for urgency.

Keywords: arsenite, arsenate, Bacillus licheniformis, utilization

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Authors: Jyosthna Khanna Goli, Shaik Naseeruddin, Hameeda Bee

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Employing lignocellulosic biomass in fermentative production of xylitol and bioethanol is gaining interest as it is renewable, cheap, and abundantly available. Xylitol is a polyol, gaining its importance in the food and pharmacological industry due to its low calorific value and anti-cariogenic nature. Bioethanol from lignocellulosic biomass is widely accepted as an alternative fuel for transportation with reduced CO₂ emissions, thus reducing the greenhouse effect. Typha latifolia, an aquatic weed, was found to be promising lignocellulosic substrate as it posses a high amount of sugars and does not compete with arable lands and interfere with food and feed competition. In the present study, xylose from hemicellulosic fraction of typha is converted to xylitol by isolate Jfh5 (Candida. tropicalis) and cellulose part to ethanol using Saccharomyces cerevisiaeVS3. Initially, alkali pretreatment of typha using sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, sodium bisulphate and sodium dithionate for overnight (18h) at room temperature (28 ± 2°C), resulted in maximum delignification of 75% with 2% (v/v) sodium bisulphate. Later, pretreated biomass was subjected to acid hydrolysis with 1%, 1.5%, 2%, and 3% H₂SO₄ at 110 °C and 121°C for 30 and 60 min, respectively. 2% H₂SO₄ at 121°C for 60 min was found to release 13.5 g /l sugars, which on detoxification and fermentation produced 8.1g/l xylitol with yield and productivity of 0.65g/g and 0.112g/l/h respectively. Further enzymatic hydrolysis of the residual substrate obtained after acid hydrolysis released 11g/l sugar, which on fermentation with VS3 produced 4.9g/l ethanol with yield and productivity of 0.22g/g and 0.136g/l/h respectively.

Keywords: delignification, xylitol, bioethanol, acid hydrolysis, enzyme hydrolysis

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Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

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Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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Authors: Aditi Taunk, George Iskander, Kitty Ka Kit Ho, Mark Willcox, Naresh Kumar

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Bacterial infections on biomaterial implants and medical devices accounts for 60-70% of all hospital acquired infections (HAIs). Treatment or removal of these infected devices results in high patient mortality and morbidity along with increased hospital expenses. In addition, with no effective strategies currently available and rapid development of antibacterial resistance has made device-related infections extremely difficult to treat. Therefore, in this project we have developed biomaterial surfaces using antibacterial compounds that inhibit biofilm formation by interfering with the bacterial communication mechanism known as quorum sensing (QS). This study focuses on covalent attachment of potent quorum sensing (QS) inhibiting compounds, halogenated furanones (FUs) and dihydropyrrol-2-ones (DHPs), onto glass surfaces. The FUs were attached by photoactivating the azide groups on the surface, and the acid functionalized DHPs were immobilized on amine surface via EDC/NHS coupling. The modified surfaces were tested in vitro against pathogenic organisms such as Staphylococcus aureus and Pseudomonas aeruginosa using confocal laser scanning microscopy (CLSM). Successful attachment of compounds on the substrates was confirmed by X-ray photoelectron spectroscopy (XPS) and contact angle measurements. The antibacterial efficacy was assessed, and significant reduction in bacterial adhesion and biofilm formation was observed on the FU and DHP coated surfaces. The activity of the coating was dependent upon the type of substituent present on the phenyl group of the DHP compound. For example, the ortho-fluorophenyl DHP (DHP-2) exhibited 79% reduction in bacterial adhesion against S. aureus and para-fluorophenyl DHP (DHP-3) exhibited 70% reduction against P. aeruginosa. The results were found to be comparable to DHP coated surfaces prepared in earlier study via Michael addition reaction. FUs and DHPs were able to retain their in vitro antibacterial efficacy after covalent attachment via azide chemistry. This approach is a promising strategy to develop efficient antibacterial biomaterials to reduce device related infections.

Keywords: antibacterial biomaterials, biomedical device-related infections, quorum sensing, surface functionalization

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Authors: Ali Bahkali, Rayan Yousef Booq, Mohammad Khiyami

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Soil samples were collected from five different regions in the Kingdom of Saudi Arabia. Microbiological methods included dilution methods and pour plates to isolate and purify bacteria soil. The ability of isolates to develop biopolymer was investigated on petri dishes containing elements and substance concentrations stimulating developing biopolymer. Fluorescent stains, Nile red and Nile blue were used to stain the bacterial cells developing biopolymers. In addition, Sudan black was used to detect biopolymers in bacterial cells. The isolates which developed biopolymers were identified based on their gene sequence of 1 6sRNA and their ability to grow and synthesize PHAs on mineral medium supplemented with 1% dates molasses as the only carbon source under nitrogen limitation. During the study 293 bacterial isolates were isolated and detected. Through the initial survey on the petri dishes, 84 isolates showed the ability to develop biopolymers. These bacterial colonies developed a pink color due to accumulation of the biopolymers in the cells. Twenty-three isolates were able to grow on dates molasses, three strains of which showed the ability to accumulate biopolymers. These strains included Bacillus sp., Ralstonia sp. and Microbacterium sp. They were detected by Nile blue A stain with fluorescence microscopy (OLYMPUS IX 51). Among the isolated strains Ralstonia sp. was selected after its ability to grow on molasses dates in the presence of a limited nitrogen source was detected. The optimum conditions for formation of biopolymers by isolated strains were investigated. Conditions studied included, best incubation duration (2 days), temperature (30°C) and pH (7-8). The maximum PHB production was raised by 1% (v1v) when using concentrations of dates molasses 1, 2, 3, 4 and 5% in MSM. The best inoculated with 1% old inoculum (1= OD). The ideal extraction method of PHA and PHB proved to be 0.4% sodium hypochlorite solution, producing a quantity of polymer 98.79% of the cell's dry weight. The maximum PHB production was 1.79 g/L recorded by Ralstonia sp. after 48 h, while it was 1.40 g/L produced by R.eutropha ATCC 17697 after 48 h.

Keywords: bacteria forming polyhydroxyalkanoate, detection, molecular, Saudi Arabia

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