Search results for: anaerobic bacteria

Authors: Dibyakanta Seth, Hari Niwas Mishra, Sankar Chandra Deka

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Microencapsulation is an established method of protecting bacteria from the adverse conditions. An improved extrusion spraying technique was used to encapsulate mixed bacteria culture of S. thermophilus and L. bulgaricus using sodium alginate as the coating material. The effect of nozzle air pressure (200, 300, 400 and 500 kPa), sodium alginate concentration (1%, 1.5%, 2%, 2.5% and 3% w/v), different concentration of calcium chloride (0.1, 0.2, 1 M) and initial cell loads (10⁷, 10⁸, 10⁹ cfu/ml) on the viability of encapsulated bacteria were investigated. With the increase in air pressure the size of microcapsules decreased, however the effect was non-significant. There was no significant difference (p > 0.05) in the viability of encapsulated cells when the concentration of calcium chloride was increased. Increased level of sodium alginate significantly increased the survival ratio of encapsulated bacteria (P < 0.01). Encapsulation with 3% alginate was treated as optimum since a higher concentration of alginate increased the gel strength of the solution and thus was difficult to spray. Under optimal conditions 3% alginate, 10⁹ cfu/ml cell load, 20 min hardening time in 0.1 M CaCl2 and 400 kPa nozzle air pressure, the viability of bacteria cells was maximum compared to the free cells. The microcapsules made at the optimal condition when mixed with yoghurt and subjected to spray drying at 148°C, the survival ratio was 2.48×10⁻¹ for S. thermophilus and 7.26×10⁻¹ for L. bulgaricus. In contrast, the survival ratio of free cells of S. thermophilus and L. bulgaricus were 2.36×10⁻³ and 8.27×10⁻³, respectively. This study showed a decline in viable cells count of about 0.5 log over a period of 7 weeks while there was a decline of about 1 log in cultures which were incorporated as free cells in yoghurt. Microencapsulation provided better protection at higher acidity compared to free cells. This study demonstrated that microencapsulation of yoghurt culture in sodium alginate is an effective technique of protection against extreme drying conditions.

Keywords: extrusion, microencapsulation, spray drying, sweetened yoghurt

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Authors: Phillip Minnaar, Neil Jolly, Louisa Beukes, Santiago Benito-Saez

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Dovyalis caffra is a tree found on the African continent. Limited information exists on the effect of acetous fermentation on the phytochemicals of Kei-apple fruit. The phytochemical content of vinegars is derived from compounds present in the fruit the vinegar is made of. Kei-apple fruit juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation (AF). Acetous fermentation followed AF, using an acetic acid bacteria consortium as an inoculant. Juice had the lowest pH and highest total acidity (TA). The wine had the highest pH and vinegars lowest TA. Total soluble solids and L-malic acid decreased during AF and acetous fermentation. Volatile acidity concentration was not different among vinegars. Gallic, syringic, caffeic, p-coumaric, and chlorogenic acids increased during acetous fermentation, whereas ferulic, sinapic, and protocatechuic acids decreased. Chlorogenic acid was the most abundant phenolic acid in both wines and vinegars. It is evident from this investigation that Kei-apple vinegar is a source of plant-derived phenolics, which evolved through fermentation. However, the AAB selection showed minimal performance with respect to VA production. Acetic acid bacteria selection for acetous fermentation should be reconsidered, and the reasons for the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, acetous fermentation, liquid chromatography, phenolic acids

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Authors: Nesrine Lenchi, Salima Kebbouche-Gana, Laddada Belaid, Mohamed Lamine Khelfaoui, Mohamed Lamine Gana

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Saline lakes are extreme hypersaline environments that are considered five to ten times saltier than seawater (150 – 300 g L-1 salt concentration). Hypersaline regions differ from each other in terms of salt concentration, chemical composition and geographical location, which determine the nature of inhabitant microorganisms. In order to explore the diversity of moderate and extreme halophiles Bacteria in Chott Tinsilt (East of Algeria), an isolation program was performed. In the first time, water samples were collected from the saltern during pre-salt harvesting phase. Salinity, pH and temperature of the sampling site were determined in situ. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions. Isolates were obtained by plating out the samples in complex and synthetic media. In this study, seven halophiles cultures of Bacteria were isolated. Isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (oxidase, catalase, nitrate reductase and urease), and optimization of growth conditions were done. The results indicated that the salinity optima varied from 50 to 250 g L-1, whereas the optimum of temperature range from 25°C to 35°C. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. The results showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Idiomarina, Halobacillus Thalassobacillus and Planococcus genera and what may represent a new bacterial genus.

Keywords: bacteria, Chott, halophilic, 16S rRNA

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Authors: Wilailak Siripornadulsil, Siriyanapat Tasaku, Jutamas Buahorm, Surasak Siripornadulsil

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The objectives of this study were to isolate LAB from various sources, dietary supplement, Thai traditional fermented food, and freshwater fish and to characterize their potential as probiotic cultures. Out of 1,558 isolates, 730 were identified as LAB based on isolation on MRS agar supplemented with a bromocresol purple indicator and CaCO3 and gram-positive, catalase and oxidase negative characteristics. Eight isolates showed the potential probiotic properties including tolerance to acid, bile salt and heat, proteolytic, amylolytic and lipolytic activities and oxalate-degrading capability. They all showed the antimicrobial activity against some Gram-negative and Gram-positive pathogenic bacteria. Based on 16S rDNA sequence analysis, they were identified as Enterococcus faecalis BT2 and MG30, Leconostoc mesenteroides SW64 and Pediococcus pentosaceous BD33, CF32, NP6, PS34 and SW5. The health beneficial effects and food safety will be further investigated and developed as a probiotic or protective culture used in Nile tilapia belly flap meat fermentation.

Keywords: probiotic, lactic acid bacteria, pathogen, protective culture

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Authors: Zahid Rehman, TorOve Leiknes

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Quorum sensing (QS) is the process by which bacteria communicate with each other through small signaling molecules, such as N-acylhomoserine lactones (AHLs). Also, certain bacteria have the ability to degrade AHL molecules by a process referred to as quorum quenching (QQ); therefore, QQ can be used to control bacterial infections and biofilm formation. In this study, we aimed to identify new species of bacteria with QQ activities. To achieve this, sediments from Red Sea were collected either in the close vicinity of Sea grass or from area with no vegetation. From these samples, we isolated 72 bacterial strains and tested their ability to degrade/inactivate AHL molecules. Chromobacterium violaceum based bioassay was used in initial screening of isolates for QQ activity. The QQ activity of the positive isolates was further confirmed and quantified by employing liquid chromatography and mass spectrometry. These analyses showed that isolated bacterial strain could degrade AHL molecules with different acyl chain length and modifications. Sequencing of 16S-rRNA genes of positive isolates revealed that they belong to three different genera. Specifically, two isolates belong to genus Erythrobacter, four to Labrenzia and one isolate belongs to Bacterioplanes. Time course experiment showed that isolate belonging to genus Erythrobacter could degrade AHLs faster than other isolates. Furthermore, these isolates were tested for their ability to inhibit formation of biofilm and degradation of 3OXO-C12 AHLs produced by P. aeruginosa PAO1. Our results showed that isolate VG12 is better at controlling biofilm formation. This aligns with the ability of VG12 to cause at least 10-fold reduction in the amount of different AHLs tested.

Keywords: quorum sensing, biofilm, quorum quenching, anti-biofouling

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Authors: Hadjer Didouh, Mohammed Hadj Melliani, Izzeddine Sameut Bouhaik

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Corrosion is a serious problem in industrial installations or metallic transport pipes. Corrosion is an interfacial process controlled by several parameters. The presence of microorganisms affects the kinetics of corrosion. This type of corrosion is often referred to as bio-corrosion or corrosion influenced by microorganisms (MIC). The action of a microorganism or a bacterium is carried out by the formation of biofilm following its attachment to the metal surface. The formation of biofilm isolates the metal surface from its environment and allows the bacteria to control the parameters of the metal/bacteria interface. Biofilm formation by sulfate-reducing bacteria (SRB) X52 steel poses substantial challenges in the oil and gas industry SONATRACH of Algeria. This research delves into the complex attachment mechanisms employed by SRB biofilm on X52 carbon steel and investigates innovative strategies for effective mitigation using biocides. The exploration commences by elucidating the underlying mechanisms facilitating SRB biofilm adhesion to X52 carbon steel, considering factors such as surface morphology, electrostatic interactions, and microbial extracellular substances. Advanced microscopy and spectroscopic techniques provide support to the attachment processes, laying the foundation for targeted mitigation strategies. The use of 100 ppm of Moringa Oleifera extract biocide as a promising approach to control and prevent SRB biofilm formation on X52 carbon steel surfaces. Green extracts undergo evaluation for their effectiveness in disrupting biofilm development while ensuring the integrity of the steel substrate. Systematic analysis is conducted on the biocide's impact on the biofilm's structural integrity, microbial viability, and overall attachment strength. This two-pronged investigation aims to deepen our comprehension of SRB biofilm dynamics and contribute to the development of effective strategies for mitigating its impact on X52 carbon steel.

Keywords: attachment, bio-corrosion, biofilm, metal/bacteria interface

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Authors: Munir Ashraf, Shagufta Riaz

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Personal protective equipment (PPE) and bioactive textiles are highly important for the health care of front line hospital workers, patients, and the general population to be safe from highly infectious diseases. This was even more critical in the wake of COVID-19 outbreak. Most of the medical textiles are inactive against various viruses and bacteria, hence there is a need to wash them frequently to avoid the spread of microorganisms. According to survey conducted by the world health organization, more than 500 million people get infected from hospitals, and more than 13 million died due to these hospitals’ acquired deadly diseases. The market available PPE are though effective against the penetration of pathogens and to kill bacteria but, they are not breathable and active against different viruses. Therefore, there was a great need to develop textiles that are not only effective against bacteria, fungi, and viruses but also are comfortable to the medical personnel and patients. In the present study, waterproof breathable, and biologically active textiles were developed using antiviral and antibacterial nanomaterials. These nanomaterials like TiO₂, ZnO, Cu, and Ag were immobilized at the surface of cotton fabric by using different silane coupling agents and electroless deposition that they retained their functionality even after 30 industrial laundering cycles. Afterwards, the treated fabrics were coated with a waterproof breathable film to prevent the permeation of liquid droplets, any particle or microorganisms greater than 80 nm. The developed cotton fabric was highly active against bacteria and viruses. The good durability of nanomaterials at the cotton surface after several industrial washing cycles makes this fabric an ideal candidate for bioactive textiles used in the medical field.

Keywords: antibacterial, antiviral, cotton, durable

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Authors: G. Amariei, K. Boltes, R. Rosal, P. Leton

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Wastewater treatment plants (WWTPs) are supposed to hold an important place in the reduction of emerging contaminants, but provide an environment that has potential for the development and/or spread of adaptation, as bacteria are continuously mixed with contaminants at sub-inhibitory concentrations. Reviewing the literature, there are little data available regarding the use of adapted bacteria forming activated sludge community for toxicity assessment, and only individual validations have been performed. Therefore, the aim of this work was to study the toxicity of Triclosan (TCS) and Ibuprofen (IBU), individually and in binary combination, on adapted activated sludge (AS). For this purpose a battery of biomarkers were assessed, involving oxidative stress and cytotoxicity responses: glutation-S-transferase (GST), catalase (CAT) and viable cells with FDA. In addition, we compared the toxic effects on adapted bacteria with unadapted bacteria, from a previous research. Adapted AS comes from three continuous-flow AS laboratory systems; two systems received IBU and TCS, individually; while the other received the binary combination, for 14 days. After adaptation, each bacterial culture condition was exposure to IBU, TCS and the combination, at 12 h. The concentration of IBU and TCS ranged 0.5-4mg/L and 0.012-0.1 mg/L, respectively. Batch toxicity experiments were performed using Oxygraph system (Hansatech), for determining the activity of CAT enzyme based on the quantification of oxygen production rate. Fluorimetric technique was applied as well, using a Fluoroskan Ascent Fl (Thermo) for determining the activity of GST enzyme, using monochlorobimane-GSH as substrate, and to the estimation of viable cell of the sludge, by fluorescence staining using Fluorescein Diacetate (FDA). For IBU adapted sludge, CAT activity it was increased at low concentration of IBU, TCS and mixture. However, increasing the concentration the behavior was different: while IBU tends to stabilize the CAT activity, TCS and the mixture decreased this one. GST activity was significantly increased by TCS and mixture. For IBU, no variations it was observed. For TCS adapted sludge, no significant variations on CAT activity it was observed. GST activity it was significant decreased for all contaminants. For mixture adapted sludge the behaviour of CAT activity it was similar to IBU adapted sludge. GST activity it was decreased at all concentration of IBU. While the presence of TCS and mixture, respectively, increased the GST activity. These findings were consistent with the viability cells evaluation, which clearly showed a variation of sludge viability. Our results suggest that, compared with unadapted bacteria, the adapted bacteria conditions plays a relevant role in the toxicity behaviour towards activated sludge communities.

Keywords: adapted sludge community, mixture, PPCPs, toxicity

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Authors: Karthick Gopalan, Selvamohan Thankiah

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Biosurfactants are lipid compounds produced by microbes, which are amphipathic molecules consisting of hydrophophic and hydrophilic domains. In the present investigation, ten bacterial strains were isolated from petroleum oil contaminated sites near petrol bunk. Oil collapsing test, haemolytic activity were used as a criteria for primary isolation of biosurfactant producing bacteria. In this study, all the bacterial strains gave positive results. Among the ten strains, two were observed as good biosurfactant producers, they utilize the diesel as a sole carbon source. Optimization of biosurfactant producing bacteria isolated from petroleum oil contaminated sites was carried out using different parameters such as, temperature (20ºC, 25ºC, 30ºC, 37ºC and 45ºC), pH (5,6,7,8 & 9) and nitrogen sources (ammonium chloride, ammonium carbonate and sodium nitrate). Biosurfactants produced by bacteria were extracted, dried and quantified. As a result of optimization of parameters the suitable values for the production of more amount of biosurfactant by the isolated bacterial species was observed as 30ºC (0.543 gm/lt) in the pH 7 (0.537 gm/lt) with ammonium nitrate (0.431 gm/lt) as sole carbon source.

Keywords: isolation and identification, biosurfactant, microorganism, bioaugmentation

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Authors: R. A. Lahmer, A. P. Williams, S. Townsend, S. Baker, D. L. Jones

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Chitosan-arginine (Ch-arg) has been proposed as an anti-microbial agent to reduce the proliferation of spoilage and pathogenic bacteria within meat products destined for human consumption. In the current experiment its use as an antimicrobial packaging material was examined. Two different concentrations of chitosan-arginine (0.05 and 0.15 % w/w) were blended into a cellulose film (Ch-arg film). When placed in contact with chicken and beef juice inoculated with a lux-marked strain of E. coli O157, the film incorporating the highest Ch-arg concentration resulted in a small reduction of E. coli O157 in chicken juice; however, there was no effect of the Ch-arg film on E. coli O157 in beef juice. The lack of observed effect in the beef juice experiment we ascribe to insufficient surface-to-surface contact between the film and the bacteria in the beef juice and the greater presence of other Ch-arg reactive components in the juice (e.g. fats, blood cells). Results suggest that, in combination with other anti microbials, Ch-arg packaging may offers some potential for limiting the growth of pathogenic bacteria in foodstuffs; however, further research is needed to enhance their anti-microbial performance.

Keywords: cross-contamination, foodborne pathogen, polymer film, shelf life

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Authors: M. N. Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin, A. K. Abdul Jalil, M. R. Salina, M. R. Maryam, M. Rosfarizan

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The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, streptomycin sulfate, extracellular proteins, antibiotics

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Authors: E. Soboleva, E. Sergachyova, S. G. Davydenko, T. V. Meledina

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Problem of food preservation is extremely important for mankind. Viscous damage ("illness") of bread results from development of Bacillus spp. bacteria. High temperature resistant spores of this microorganism are steady against 120°C) and remain in bread during pastries, potentially causing spoilage of the final product. Scientists are interested in further characterization of bread spoiling Bacillus spp. species. Our aim was to find weather yeast Saccharomyces cerevisiae strains that are able to produce natural antimicrobial killer factor can preserve bread illness. By diffusion method, we showed yeast antagonistic activity against spore-forming bacteria. Experimental technological parameters were the same as for bakers' yeasts production on the industrial scale. Risograph test during dough fermentation demonstrated gas production. The major finding of the study was a clear indication of the presence of killer yeast strain antagonistic activity against rope in bread causing bacteria. After demonstrating antagonistic effect of S. cerevisiae on bacteria using solid nutrient medium, we tested baked bread under provocative conditions. We also measured formation of carbon dioxide in the dough, dough-making duration and quality of the final products, when using different strains of S. cerevisiae. It is determined that the use of yeast S. cerevisiae RCAM 01730 killer strain inhibits appearance of rope in bread. Thus, natural yeast antimicrobial killer toxin, produced by some S. cerevisiae strains is an anti-rope in bread protector.

Keywords: bakers' yeasts, killer toxin, rope in bread, Saccharomyces cerevisiæ

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Authors: Yong Geun Yun, Jong Hui kim, Sang Ho Baik

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This study was carried out to develop a fermented dandelion (Tarazacum officinale) beverage using lactic acid bacteria with cinnamoyl esterase (CE) activity isolated from human feces. Lactic acid bacteria were screened based on bacterial survival ability in dandelion extract and CE activity. Dandelion extract fermented by Lactobacillus acidophilus F-46 (LA-F46) maintained approximately 105-106 log CFU/mL over an 8 days period. After fermented dandelion beverage (FDB) with LA-46 for 8 days at 37oC the pH was decreased from pH 7.0 to 3.5. Antioxidant activity by using DPPH radical scavenging activity of the prepared FDB was significantly increased compared to that of non-fermented dandelion beverage (NFDB). Moreover, CE activity was significantly enhanced during fermentation and showed the approximately 4.3 times increased concentration of caffeic acid up to 9.91 mg/100 mL after 8 days of incubation compared to NFDB. Therefore, it concluded that dandelion can be a good source for preparing a functional beverage and fermentation by LA-F46 enhanced the food functionality with enhanced caffeic acids.

Keywords: cinnamoyl esterase, dandelion, fermented beverage, lactic acid bacteria

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Authors: Elizabeth G. Hereford, Geoffrey W. Gearner

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Logan’s Branch, within the Triplett Creek Watershed of Rowan County, Kentucky, is a waterway located near important agricultural and residential areas. Part of Logan’s Branch flows over an exposed black shale formation with elevated radioactivity and heavy metals. Three sites were chosen in relation to the formation and sampled five times over a thirty-day period during the recreational season. A fourth site in North Fork in Rowan County, Kentucky was also sampled periodically as it too has contact with the shale formation. These sites were then sampled monthly. All samples are analyzed for concentrations of Escherichia coli, heterotrophic bacteria, and total coliform bacteria utilizing the membrane filtration method and various culture media. Current data suggests that the radioactivity of the shale formation influences the bacteriological growth present in the waterway; however, further data will be collected and compared with that of my colleagues to confirm this trend.

Keywords: bacteriological analysis, Escherichia coli, heterotrophic bacteria, radioactive black shale formation, water quality

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Authors: Lotem Sarid, Meirav Trebicz-Geffen, Serge Ankri

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Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the nucleobase of queuosine, they must obtain it from their diet or gut microbiota. Our previous work investigated the effects of queuine on the physiology of the eukaryotic parasite Entamoeba histolytica and defined the enzyme EhTGT responsible for its incorporation into tRNA. To our best knowledge, it is unknown how E. histolytica salvages Q from gut bacteria. We used N-acryloyl-3-aminophenylboronic acid (APB) PAGE analysis to demonstrate that E. histolytica trophozoites can salvage queuine from Q or E. coli K12 but not from the modified E. coli QueC strain, which cannot produce queuine. Next, we examined the role of EhDUF2419, a protein with homology to DNA glycosylase, as a queuine salvage enzyme in E. histolytica. When EhDUF2419 expression is silenced, it inhibits Q's conversion to queuine, resulting in a decrease in Q-tRNA levels. We also observed that Q protects control trophozoites from oxidative stress (OS), but not siEhDUF2419 trophozoites. Overall, our data reveal that EhDUF2419 is central for the salvaging of queuine from bacteria and for the resistance of the parasite to OS.

Keywords: entamoeba histolytica, epitranscriptomics, gut microbiota, queuine, queuosine, response to oxidative stress, tRNA modification.

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Authors: Vojtech Stejskal, Hana Skalova, Petr Kvapil, George Hutchinson

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Algae growth has been an important issue in water management of water plants, ponds and lakes, swimming pools, aquaculture & fish farms, gardens or golf courses for last decades. There are solutions based on chemical or biological principles. Apart of these traditional principles for inhibition of algae growth and biofilm production there are also physical methods which are very competitive compared to the traditional ones. Ultrasonic technology is one of these alternatives. Ultrasonic emitter is able to eliminate the biofilm which behaves as a host and attachment point for algae and is original reason for the algae growth. The ultrasound waves prevent majority of the bacteria in planktonic form becoming strongly attached sessile bacteria that creates welcoming layer for the biofilm production. Biofilm creation is very fast – in the serene water it takes between 30 minutes to 4 hours, depending on temperature and other parameters. Ultrasound device is not killing bacteria. Ultrasound waves are passing through bacteria, which retract as if they were in very turbulent water even though the water is visually completely serene. In these conditions, bacteria does not excrete the polysaccharide glue they use to attach to the surface of the pool or pond, where ultrasonic technology is used. Ultrasonic waves decrease the production of biofilm on the surfaces in the selected area. In case there are already at the start of the application of ultrasonic technology in a pond or basin clean inner surfaces, the biofilm production is almost absolutely inhibited. This paper talks about two different pilot applications – one in Czech Republic and second in United States of America, where the used ultrasonic technology (AlgaeControl) is coming from. On both sites, there was used Mezzo Ultrasonic Algae Control System with very positive results not only on biofilm production, but also algae growth in the surrounding area. Technology has been successfully tested in two different environments. The poster describes the differences and their influence on the efficiency of ultrasonic technology application. Conclusions and lessons learned can be possibly applied also on other sites within Europe or even further.

Keywords: algae growth, biofilm production, ultrasonic solution, ultrasound

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Authors: Khamdan Cahyari, Gumbolo Hadi Susanto, Pratikno Hidayat, Sukirman

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University canteen waste was used as raw material to produce biogas in Faculty of Industrial Technology, Islamic University of Indonesia. This faculty was home to more than 3000 students and lecturers who work and study for 5 days/week (8 hours/day). It produced approximately 85 ton/year organic fraction of canteen waste. Yet, this waste had been dumped for years in landfill area which cause severe environmental problems. It was proposed to utilize the waste as raw material for producing renewable energy source of biogas. This research activities was meant to investigate the effect of organic loading rate (OLR) and retention time (RT) of continuous anaerobic digestion process for 200 days. Organic loading rate was set at value 2, 3, 4 and 5 g VS/l/d whereas the retention time was adjusted at 30, 24, 18 and 14.4 days. Optimum condition was achieved at OLR 4 g VS/l/d and RT 24 days with biogas production rate between 0.75 to 1.25 liter/day (40-60% CH4). This indicated that the utilization of canteen waste to produce biogas was promising method to mitigate environmental problem of university canteen waste. Furthermore, biogas could be used as alternative energy source to supply energy demand at the university. This implementation is simultaneous solution for both waste and energy problems to achieve green campus.

Keywords: canteen waste, biogas, anaerobic digestion, university, green campus

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Authors: Karan Bhandari

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Almost quarter region of Indian terrain is covered by grasslands. Grass being a low maintenance perennial crop is in abundance. Farmers are well acquainted with its nature, yield and storage. The aim of this paper is to study and identify the applicability of grass as a source of bio fuel. Anaerobic break down is a well-recognized technology. This process is vital for harnessing bio fuel from grass. Grass is a lignocellulosic material which is fibrous and can readily cause problems with parts in motion. Further, it also has a tendency to float. This paper also deals with the ideal digester configuration for biogas generation from grass. Intensive analysis of the literature is studied on the optimum production of grass storage in accordance with bio digester specifications. Subsequent to this two different digester systems were designed, fabricated, analyzed. The first setup was a double stage wet continuous arrangement usually known as a Continuously Stirred Tank Reactor (CSTR). The next was a double stage, double phase system implementing Sequentially Fed Leach Beds using an Upflow Anaerobic Sludge Blanket (SLBR-UASB). The above methodologies were carried for the same feedstock acquired from the same field. Examination of grass silage was undertaken using Biomethane Potential values. The outcomes portrayed that the Continuously Stirred Tank Reactor system produced about 450 liters of methane per Kg of volatile solids, at a detention period of 48 days. The second method involving Leach Beds produced about 340 liters of methane per Kg of volatile solids with a detention period of 28 days. The results showcased that CSTR when designed exclusively for grass proved to be extremely efficient in methane production. The SLBR-UASB has significant potential to allow for lower detention times with significant levels of methane production. This technology has immense future for research and development in India in terms utilizing of grass crop as a non-conventional source of fuel.

Keywords: biomethane potential values, bio digester specifications, continuously stirred tank reactor, upflow anaerobic sludge blanket

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Authors: Fatima M. Kabbashi, Abdalla M. Abdalla, Hussam K. Hamad, Elias S. Hassan

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This study investigates the production of renewable energy (biogas) from biomedical hazard waste (blood) and eco-friendly disposal. Biogas is produced by the bacterial anaerobic digestion of biomaterial (blood). During digestion process bacterial feeding result in breaking down chemical bonds of the biomaterial and changing its features, by the end of the digestion (biogas production) the remains become manure as known. That has led to the economic and eco-friendly disposal of hazard biomedical waste (blood). The samples (Whole blood, Red blood cells 'RBCs', Blood platelet and Fresh Frozen Plasma ‘FFP’) are collected and measured in terms of carbon to nitrogen C/N ratio and total solid, then filled in connected flasks (three flasks) using water displacement method. The results of trails showed that the platelet and FFP failed to produce flammable gas, but via a gas analyzer, it showed the presence of the following gases: CO, HC, CO₂, and NOX. Otherwise, the blood and RBCs produced flammable gases: Methane-nitrous CH₃NO (99.45%), which has a blue color flame and carbon dioxide CO₂ (0.55%), which has red/yellow color flame. Methane-nitrous is sometimes used as fuel for rockets, some aircraft and racing cars.

Keywords: renewable energy, biogas, biomedical waste, blood, anaerobic digestion, eco-friendly disposal

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Authors: Aliasghar Golmohammadian, Sona Rostampour Yasouri

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One of the most important intestinal pathogenic strains is E. coli O₁₅₇:H₇. This pathogenic bacterium is transmitted to humans through water and food. E. coli O₁₅₇:H₇ is the main cause of Hemorrhagic colitis (HC), Hemolytic Uremic Syndrome (HUS), Thrombotic Thrombocytopenic Purpura (TTP) and in some cases death. Since E. coli O₁₅₇:H₇ can be transmitted through the consumption of different foods, including vegetables, agricultural products, and fresh dairy products, this study aims to identify and isolate E. coli O₁₅₇:H₇ from wastewater by PCR technique. One hundred twenty samples of water and wastewater were collected by Falcom Sterile from Shahrood and Neka cities. The samples were checked for colony formation after appropriate centrifugation and cultivation in the specific medium of Sorbitol MacConkey Agar (SMAC) and other diagnostic media of E. coli O₁₅₇:H₇. Also, the plates were observed macroscopically and microscopically. Then, the necessary phenotypic tests were performed on the colonies, and finally, after DNA extraction, the PCR technique was performed with specific primers related to rfbE and stx2 genes. The number of 5 samples (6%) out of all the samples examined were determined positive by PCR technique with observing the bands related to the mentioned genes on the agarose gel electrophoresis. PCR is a fast and accurate method to identify the bacteria E. coli O₁₅₇:H₇. Considering that E. coli bacteria is a resistant bacteria and survives in water and food for weeks and months, the PCR technique can provide the possibility of quick detection of contaminated water. Moreover, it helps people in the community control and prevent the transfer of bacteria to healthy and underground water and agricultural and even dairy products.

Keywords: E. coli O₁₅₇:H₇, PCR, water, wastewater

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Authors: Zhihao Peng, Qiong Zhang, Xiyao Li, Yongzhen Peng

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Nowadays, conventional nitrogen removal process (nitrification and denitrification) was adopted in most wastewater treatment plants, but many problems have occurred, such as: high aeration energy consumption, extra carbon sources dosage and high sludge treatment costs. The emergence of anammox has bring about the great revolution to the nitrogen removal technology, and only the ammonia and nitrite were required to remove nitrogen autotrophically, no demand for aeration and sludge treatment. However, there existed many challenges in anammox applications: difficulty of biomass retention, insufficiency of nitrite substrate, damage from complex organic etc. Much effort was put into the research in overcoming the above challenges, and the payment was rewarded. It was also imperative to establish an innovative process that can settle the above problems synchronously, after all any obstacle above mentioned can cause the collapse of anammox system. Therefore, in this study, a two-stage process was established that the sequencing batch reactor (SBR) and upflow anaerobic sludge blanket (UASB) were used in the pre-stage and post-stage, respectively. The domestic wastewater entered into the SBR first and went through anaerobic/aerobic/anoxic (An/O/A) mode, and the draining at the aerobic end of SBR was mixed with domestic wastewater, the mixture then entering to the UASB. In the long term, organic and nitrogen removal performance was evaluated. All along the operation, most COD was removed in pre-stage (COD removal efficiency > 64.1%), including some macromolecular organic matter, like: tryptophan, tyrosinase and fulvic acid, which could weaken the damage of organic matter to anammox. And the An/O/A operating mode of SBR was beneficial to the achievement and maintenance of partial nitrification (PN). Hence, sufficient and steady nitrite supply was another favorable condition to anammox enhancement. Besides, the flexible mixing ratio helped to gain a substrate ratio appropriate to anammox (1.32-1.46), which further enhance the anammox. Further, the UASB was used and gas recirculation strategy was adopted in the post-stage, aiming to achieve granulation by the selection pressure. As expected, the granules formed rapidly during 38 days, which increased from 153.3 to 354.3 μm. Based on bioactivity and gene measurement, the anammox metabolism and abundance level rose evidently, by 2.35 mgN/gVss·h and 5.3 x109. The anammox bacteria mainly distributed in the large granules (>1000 μm), while the biomass in the flocs (<200 μm) and microgranules (200-500 μm) barely displayed anammox bioactivity. Enhanced anammox promoted the advanced autotrophic nitrogen removal, which increased from 71.9% to 93.4%, even when the temperature was only 12.9 ℃. Therefore, it was feasible to enhance anammox in the multiple favorable conditions created, and the strategy extended the application of anammox to the full-scale mainstream, enhanced the understanding of anammox in the aspects of culturing conditions.

Keywords: anammox, granules, nitrite accumulation, nitrogen removal efficiency

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Authors: Ahmed Elreedy, Ahmed Tawfik

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This investigation aims to assess the effect of inoculum to substrate ratio (ISR) and nitrogen to phosphorous balance on simultaneous biohydrogen and methane production from anaerobic decomposition of mono-ethylene glycol (MEG). Different ISRs were applied in the range between 2.65 and 13.23 gVSS/gCOD, whereas the tested N/P ratios were changed from 4.6 to 8.5; both under thermophilic conditions (55°C). The maximum obtained methane and hydrogen yields (MY and HY) of 151.86±10.8 and 22.27±1.1 mL/gCODinitial were recorded at ISRs of 5.29 and 3.78 gVSS/gCOD, respectively. Unlikely, the ammonification process, in terms of net ammonia produced, was found to be ISR and COD/N ratio dependent, reaching its peak value of 515.5±31.05 mgNH4-N/L at ISR and COD/N ratio of 13.23 gVSS/gCOD and 11.56. The optimum HY was enhanced by more than 1.45-fold with declining N/P ratio from 8.5 to 4.6; whereas, the MY was improved (1.6-fold), while increasing N/P ratio from 4.6 to 5.5 with no significant impact at N/P ratio of 8.5. The results obtained revealed that the methane production was strongly influenced by initial ammonia, compared to initial phosphate. Likewise, the generation of ammonia was markedly deteriorated from 535.25±41.5 to 238.33±17.6 mgNH4-N/L with increasing N/P ratio from 4.6 to 8.5. The kinetic study using Modified Gompertz equation was successfully fitted to the experimental outputs (R2 > 0.9761).

Keywords: mono-ethylene glycol, biohydrogen and methane, inoculum to substrate ratio, nitrogen to phosphorous balance, ammonification

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Authors: H. Nabahat, E. Amiri, F. AzaditalabDavoudabadi, N. Zaeri

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Tooth decay is one of the most common forms of oral and dental illness in the world, which causes huge costs of treatment, especially in high-risk groups such as people with oral dry mouth, prevention and control of it are very important. The use of traditional treatments such as extraction of drugs from medicinal plants is of paramount importance to Iran and the international community as well. The present study was conducted with the aim of investigating the antibacterial effect of the extract of Salvia officinalis and Mentha pulegium, which are the most commonly used drugs in the treatment of oral and teeth bacterial (Streptococcus mutant, Lactobacillus rhamnosis, and Actinomyces viscosis) in vitro method. In this experimental study, two herbs of Salvia and Mentha were prepared by maceration of hydroalcoholic extract, and the antibacterial effect was evaluated by broth macro dilution on streptococcal mutagen bacteria, lactobacillus rhamnosis, and viscose actinomycosis. The results were analyzed by the Whitney Mann test (P > 0.05). The results showed that the minimum inhibitory concentration (MIC) of the salmonella extract for Streptococcus mutan were 6.25 and 12.5 μg/ml, respectively, for lactobacillus of 1.56 and 3.12 μg/ml, respectively, and for actinomycosis viscose, The order of 12.5 and 100 μg/ml was obtained. As a result, broth macro dilution showed that both extracts of Salvia and Mentha had an inhibitory effect on all three species of bacteria. This effect for Salvia was significantly (P < 0.05) more than Mentha and was within the concentration range of both the extracts and had a bactericidal effect on all three bacteria.

Keywords: antibacterial effect, dental bacteria, herbal extracts , salvia officinalis, mentha pulegium

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Authors: Aastha Paliwal, H. N. Chanakya, S. Dasappa

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Lignocellulose, as an alternate feedstock for biogas production, has been an active area of research. However, lignocellulose poses a lot of operational difficulties- widespread variation in the structural organization of lignocellulosic matrix, amenability to degradation, low bulk density, to name a few. Amongst these, the low bulk density of the lignocellulosic feedstock is crucial to the process operation and optimization. Low bulk densities render the feedstock floating in conventional liquid/wet digesters. Low bulk densities also restrict the maximum achievable organic loading rate in the reactor, decreasing the power density of the reactor. However, during digestion, lignocellulose undergoes very high compaction (up to 26 times feeding density). This first reduces the achievable OLR (because of low feeding density) and compaction during digestion, then renders the reactor space underutilized and also imposes significant mass transfer limitations. The objective of this paper was to understand the effects of compacting lignocellulose on mass transfer and the influence of loss of different components on the bulk density and hence structural integrity of the digesting lignocellulosic feedstock. 10 different lignocellulosic feedstocks (monocots and dicots) were digested anaerobically in a fed-batch, leach bed reactor -solid-state stratified bed reactor (SSBR). Percolation rates of the recycled bio-digester liquid (BDL) were also measured during the reactor run period to understand the implication of compaction on mass transfer. After 95 ds, in a destructive sampling, lignocellulosic feedstocks digested at different SRT were investigated to quantitate the weekly changes in bulk density and lignocellulosic composition. Further, percolation rate data was also compared to bulk density data. Results from the study indicate loss of hemicellulose (r²=0.76), hot water extractives (r²=0.68), and oxalate extractives (r²=0.64) had dominant influence on changing the structural integrity of the studied lignocellulose during anaerobic digestion. Further, feeding bulk density of the lignocellulose can be maintained between 300-400kg/m³ to achieve higher OLR, and bulk density of 440-500kg/m³ incurs significant mass transfer limitation for high compacting beds of dicots.

Keywords: anaerobic digestion, bulk density, feed compaction, lignocellulose, lignocellulosic matrix, cellulose, hemicellulose, lignin, extractives, mass transfer

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Authors: Pavel Bulai, Taras Pitlik, Tatsiana Kulahava, Timofei Lipski

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The method of Electrocardiography (ECG) interpretation for post-exercise recovery tracking was developed. Metabolic indices (aerobic and anaerobic) were designed using ECG-derived features. This study reports the associations between aerobic and anaerobic indices and classical parameters of the person’s physiological state, including blood biochemistry, glycogen concentration and VO2max changes. During the study 9 participants, healthy, physically active medium trained men and women, which trained 2-4 times per week for at least 9 weeks, fulfilled (i) ECG monitoring using Apple Watch Series 4 (AWS4); (ii) blood biochemical analysis; (iii) maximal oxygen consumption (VO2max) test, (iv) bioimpedance analysis (BIA). ECG signals from a single-lead wrist-wearable device were processed with detection of QRS-complex. Aerobic index (AI) was derived as the normalized slope of QR segment. Anaerobic index (ANI) was derived as the normalized slope of SJ segment. Biochemical parameters, glycogen content and VO2max were evaluated eight times within 3-60 hours after training. ECGs were recorded 5 times per day, plus before and after training, cycloergometry and BIA. The negative correlation between AI and blood markers of the muscles functional status including creatine phosphokinase (r=-0.238, p < 0.008), aspartate aminotransferase (r=-0.249, p < 0.004) and uric acid (r = -0.293, p<0.004) were observed. ANI was also correlated with creatine phosphokinase (r= -0.265, p < 0.003), aspartate aminotransferase (r = -0.292, p < 0.001), lactate dehydrogenase (LDH) (r = -0.190, p < 0.050). So, when the level of muscular enzymes increases during post-exercise fatigue, AI and ANI decrease. During recovery, the level of metabolites is restored, and metabolic indices rising is registered. It can be concluded that AI and ANI adequately reflect the physiology of the muscles during recovery. One of the markers of an athlete’s physiological state is the ratio between testosterone and cortisol (TCR). TCR provides a relative indication of anabolic-catabolic balance and is considered to be more sensitive to training stress than measuring testosterone and cortisol separately. AI shows a strong negative correlation with TCR (r=-0.437, p < 0.001) and correctly represents post-exercise physiology. In order to reveal the relation between the ECG-derived metabolic indices and the state of the cardiorespiratory system, direct measurements of VO2max were carried out at various time points after training sessions. The negative correlation between AI and VO2max (r = -0.342, p < 0.001) was obtained. These data testifying VO2max rising during fatigue are controversial. However, some studies have revealed increased stroke volume after training, that agrees with findings. It is important to note that post-exercise increase in VO2max does not mean an athlete’s readiness for the next training session, because the recovery of the cardiovascular system occurs over a substantially longer period. Negative correlations registered for ANI with glycogen (r = -0.303, p < 0.001), albumin (r = -0.205, p < 0.021) and creatinine (r = -0.268, p < 0.002) reflect the dehydration status of participants after training. Correlations between designed metabolic indices and physiological parameters revealed in this study can be considered as the sufficient evidence to use these indices for assessing the state of person’s aerobic and anaerobic metabolic systems after training during fatigue, recovery and supercompensation.

Keywords: aerobic index, anaerobic index, electrocardiography, supercompensation

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Authors: B. Tayebani, N. Hosseinibalam, D. Mostofinejad

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Interest in using bacteria in cement materials due to its positive influences has been increased. Cement materials such as mortar and concrete basically suffer from higher porosity and water absorption compared to other building materials such as steel materials. Because of the negative side-effects of certain chemical techniques, biological methods have been proposed as a desired and environmentally friendly strategy for reducing concrete porosity and diminishing water absorption. This paper presents the results of an experimental investigation carried out to evaluate the influence of Sporosarcina pasteurii bacteria on the behaviour of two types of concretes (light weight aggregate concrete and normal weight concrete). The resistance of specimens to water penetration by testing water absorption and evaluating the electrical resistance of those concretes was examined and compared. As a conclusion, 20% increase in electrical resistance and 10% reduction in water absorption of lightweight aggregate concrete (LWAC) and for normal concrete the results show 7% decrease in water absorption and almost 10% increase in electrical resistance.

Keywords: bacteria, biological method, normal weight concrete, lightweight aggregate concrete, water absorption, electrical resistance

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Authors: Negar Zaheddoust, Negin Zaheddoust, Abbas Asoudeh-Fard

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Probiotic bacteria have been employed as a novel and less side-effect strategy for anticancer therapy. Since the oral cavity is a host for probiotic and pathogen bacteria to colonize, more investigation is needed to evaluate the effectiveness of this novel adjunctive treatment for oral cancer. We considered Lactobacillus plantarum as a probiotic and Streptococcus mutans as a pathogen bacterium in our study. The aim of this study is to examine the effect of Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT / PTEN, and MAPK signaling pathway, which is involved in apoptosis or survival of oral cancer KB cells. On the other hand, to study the effects of these bacteria on normal cells, we used HUVEC cells. The KB and HUVEC cell lines were co-cultured with Lactobacillus plantarum and Streptococcus mutans isolated from traditional Iranian dairy and dental plaque, respectively. The growth-inhibitory effects of these two bacteria on KB and HUVEC cells were determined by (3-(4, 5-dimethylthiazolyl-2)-2,5diphenyltetrazolium bromide) MTT assay. MTT results demonstrated that the proliferation of KB cells was affected in a time, dose, and strain-dependent manner. In the following, the examination of induced apoptosis or necrosis in co-cultured KB cells with the best IC50 concentration of the Lactobacillus plantarum and Streptococcus mutans will be analyzed by FACS flow cytometry, and the changes in gene expression of Casp3, AKT / PTEN, MAPK genes will be evaluated using real-time polymerase chain reaction.

Keywords: cancer therapy, induced apoptosis, oral cancer, probiotics

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Authors: Tsige Reda

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Essential oil of Thymus vulgaris was extracted by means of hydro-distillation. This study was done to investigate the chemical composition, antibacterial and antioxidant activities. The chemical composition of the essential oils was determined using gas chromatography coupled to mass spectroscopy (GC-MS). Using disc diffusion assay the antibacterial activity was assessed on one Gram-positive bacteria and one Gram-negative bacteria. The percentage oil yield of the essential oil was found to be 0.97 ± 0.08% (w/w) with yellow color. The physicochemical constants of the oil were also noted. The phytochemical screening of the plant extract revealed the presence of tannins, saponins, phenol, flavonoids, terpenoids, steroids and alkaloids. A total of 18 chemical constituents were identified by Gas Chromatography-Mass Spectroscopy analysis representing 100% of the total essential oil of Thymus vulgaris, with thymol (31.977%), o-cymene (29.992%), and carvacrol (14.541%). Previous studies have revealed that the thymol, o-cymen and carvacrol components of Thymus vulgaris are responsible for their biological activities. Thymus vulgaris have been used traditionally to treat a wide variety of infections. Based on the extensive use and lack of scientific evidence, a study was embarked upon to determine its bioactivity. The essential oil of Thymus vulgaris leaves exhibited higher activity towards the Gram-positive bacteria (Staphylococcus aurous) than the Gram-negative bacteria (Escherichia coli) and also has good antioxidant activity, and can be used medicinal and therapeutic applications. This activity may be due to the high amount of thymol, o-cymen and carvacrol.

Keywords: hydro-distillation, Thymus vulgaris, essential oil composition, phytochemical screening, physicochemical constants, antioxidant activity, antibacterial activity

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Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

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Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: gold nanoparticles, Gomphrena celesioides, Prunus amygdalus, pathogens

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Authors: Abheyjit Singh, Kulwant Singh

Abstract:

There is a gradual depletion of the water table in the earth's crust, and it is required to converse and reduce the scarcity of water. This is only done by rainwater harvesting, recycling of water and by judicially consumption/utilization of water and adopting unique treatment measures. Domestic waste is generated in residential areas, commercial settings, and institutions. Waste, in general, is unwanted, undesirable, and nevertheless an inevitable and inherent product of social, economic, and cultural life. In a cluster, a need-based system is formed where the project is designed for systematic analysis, collection of sewage from the cluster, treating it and then recycling it for multifarious work. The liquid waste may consist of Sanitary sewage/ Domestic waste, Industrial waste, Storm waste, or Mixed Waste. The sewage contains both suspended and dissolved particles, and the total amount of organic material is related to the strength of the sewage. The untreated domestic sanitary sewage has a BOD (Biochemical Oxygen Demand) of 200 mg/l. TSS (Total Suspended Solids) about 240 mg/l. Industrial Waste may have BOD and TSS values much higher than those of sanitary sewage. Another type of impurities of wastewater is plant nutrients, especially when there are compounds of nitrogen N phosphorus P in the sewage; raw sanitary contains approx. 35 mg/l Nitrogen and 10 mg/l of Phosphorus. Finally, the pathogen in the waste is expected to be proportional to the concentration of facial coliform bacteria. The coliform concentration in raw sanitary sewage is roughly 1 billion per liter. The system of sewage disposal technique has been universally applied to all conditions, which are the nature of soil formation, Availability of land, Quantity of Sewage to be disposed of, The degree of treatment and the relative cost of disposal technique. The adopted Thappar Model (India) has the following designed parameters consisting of a Screen Chamber, a Digestion Tank, a Skimming Tank, a Stabilization Tank, an Oxidation Pond and a Water Storage Pond. The screening Chamber is used to remove plastic and other solids, The Digestion Tank is designed as an anaerobic tank having a retention period of 8 hours, The Skimming Tank has an outlet that is kept 1 meter below the surface anaerobic condition at the bottom and also help in organic solid remover, Stabilization Tank is designed as primary settling tank, Oxidation Pond is a facultative pond having a depth of 1.5 meter, Storage Pond is designed as per the requirement. The cost of the Thappar model is Rs. 185 Lakh per 3,000 to 4,000 population, and the Area required is 1.5 Acre. The complete structure will linning as per the requirement. The annual maintenance will be Rs. 5 lakh per year. The project is useful for water conservation, silage water for irrigation, decrease of BOD and there will be no longer damage to community assets and economic loss to the farmer community by inundation. There will be a healthy and clean environment in the community.

Keywords: collection, treatment, utilization, economic

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