Search results for: bacterial chemotaxis

Authors: P. M. G. Pathiraja, P. Egodawatta, A. Goonetilleke, V. S. J. Te'o

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The production and use of Polychlorinated biphenyls (PCBs), a group of synthetic halogenated hydrocarbons have been restricted worldwide due to its toxicity and categorized as one of the twelve priority persistent organic pollutants (POP) by the Stockholm Convention. Low reactivity and high chemical stability of PCBs have made them highly persistent in the environment and bio-concentration and bio-magnification along the food chain contribute to multiple health impacts in humans and animals. Remediating environments contaminated with PCBs is a challenging task for decades. Use of microorganisms for remediation of PCB contaminated soils and sediments have been widely investigated due to the potential of breakdown these complex contaminants with minimum environmental impacts. To achieve an effective bioremediation of polychlorinated biphenyls (PCBs) contaminated environments, microbes were sourced from environmental samples and tested for their ability to hydrolyze PCBs under different conditions. Comparison of PCB degradation efficiencies of four naturally occurring facultative bacterial cultures isolated through selective enrichment under aerobic and anaerobic conditions were simultaneously investigated in minimal salt medium using 50 mg/L Aroclor 1260, a commonly used commercial PCB mixture as the sole source of carbon. The results of a six-week study demonstrated that all the tested facultative Achromobacter, Ochrobactrum, Lysinibacillus and Pseudomonas strains are capable of degrading PCBs under both anaerobic and aerobic conditions while assisting hydrophobic PCBs to make solubilize in the aqueous minimal medium. Overall, the results suggest that some facultative bacteria are capable of effective in degrading PCBs under anaerobic conditions through reductive dechlorination and under aerobic conditions through oxidation. Therefore, use of suitable facultative microorganisms under combined anaerobic-aerobic conditions and combination of such strains capable of solubilization and breakdown of PCBs has high potential in achieving higher PCB removal rates.

Keywords: bioremediation, combined anaerobic-aerobic degradation, facultative microorganisms, polychlorinated biphenyls

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Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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Authors: Albert P. Ekanem, Austin I. Obiekezie, Elizabeth X. Ntia

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Introduction: Some African plants and bile from animals have shown efficacies in the treatment and control of diseases in farmed fish. The background of the study is based on the fact the African rain forest is blessed with abundance of medicinal plants that should be investigated for their use in the treatment of diseases. The significance of the study is informed by the fact that chemical-based substances accumulates in the tissues of food fish, thereby reducing the food values of such products and moreover, the continuous use of chemotherapeutants in the aquatic environments tends to degrades the affected environment. Methodology: Plants and animal products were extracted, purified and applied under in vitro and in vivo conditions to the affected organisms. Effective plants and biles were analyzed for active biological substances responsible for the activities by both qualitative and HPLC methods. Results: Extracts of Carica papaya and Mucuna pruriens were effective in the treatment of Ichthyophthiriasis in goldfish (Carassius auratus auratus) with high host tolerance. Similarly, ectoparasitic monogeneans were effectively dislodged from the gills and skin of goldfish by the application of extracts of Piper guineense at therapeutic concentrations. Artemesia annua with known antimalarial activities in human was also effective against fish monogenean parasites of Clarias gariepinus in a concentration related manner without detriments to the host. Effective antibacterial activities against Aeromonas and Pseudomonas diseases of the African catfish (Heterobranchus longifilis) were demonstrated in some plants such as Phylanthus amarus, Allium sativum, A. annua, and Citrus lemon. Bile from some animals (fish, goat, chicken, cow, and pig) showed great antibacterial activities against some gastrointestinal bacterial pathogens of fish. Conclusions: African plants and some animal bile have shown potential promise in the treatment of diseases in fish and other aquatic animals. The use of chemical-based substances for control of diseases in the aquatic environments should be restricted.

Keywords: control, diseases, fish, natural products, treatment

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Authors: Inayat Ur Rahman Arshad

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Stevia rebaudiana B. is a shrubby perennial herb of Asteraceae family that possesses the unique ability of accumulative non caloric sweet Steviol Glycosides (SGs). The purpose of the study is to optimize sugar concentration, pH level and inoculum size for inducing the callus with optimum growth and efficient antibacterial potential. Three different experiments were conducted in which Callus explant from three-months-old already established callus of Stevia reabudiana of four different sizes were inoculated on Murashige and Skoog (MS) basal medium supplemented with five different sucrose concentration and pH adjusted at four different levels. Maximum callus induction 100, 87.5 and 85.33% was resulted in the medium supplemented with 30g/l sucrose, pH maintained at 5.5 and inoculated with 1.25g inoculum respectively. Similarly, the highest fresh weight 65.00, 75.50 and 50.53g/l were noted in medium fortified with 40g/l sucrose, inoculated 1.25g inoculum and 6.0 pH level respectively. However, the callus developed in medium containing 50g/l sucrose found highly antibacterial potent with 27.3 and 26.5mm inhibition zone against P. vulgaris and B. subtilize respectively. Similarly, the callus grown on medium inoculated with 1.00g inoculum resulted in maximum antibacterial potential against S. aureus and P. vulgaris with 25 and 23.72mm inhibition zones respectively. However, in the case of pH levels the medium maintained at 6.5pH showed maximum antibacterial activity against P. vulgaris, B.subtilis and E.coli with 27.9, 25 and 23.72mm respectively. The ethyl acetate extract of Stevia callus and leaves did not show antibacterial potential against Xanthomonas campestris and Clavebactor michiganensis. In the entire experiment the standard antibacterial agent Streptomycin showed the highest inhibition zones from the rest of the callus extract, however the pure DMSO (Dimethyl Sulfoxide) caused no inhibitory zone against any bacteria. From these findings it is concluded that among various levels sucrose at the rate of 40g L-1, pH 6.0 and inoculums 0.75g was found best for most of the growth and quality attributes including fresh weight, dry weight and antibacterial activities and therefore can be recommended for callus proliferation and antibacterial potential of Stevia rebaudiana

Keywords: Steviol Glycosides, Skoog, Murashige, Clavebactor michiganensis

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Authors: Inayat Ur Rahman Arshad

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Background: Stevia rebaudiana B. is a shrubby perennial herb of Asteraceae family that possesses the unique ability of accumulative non-caloric sweet steviol glycosides (SGs). Purpose: The purpose of the study is to optimize sugar concentration, pH level, and inoculum size for inducing the callus with optimum growth and efficient antibacterial potential. Method: Three different experiments were conducted in which Callus explant from three-months-old already established callus of Stevia reabudiana of four different sizes was inoculated on Murashige and Skoog (MS) basal medium supplemented with five different sucrose concentration and pH adjusted at four different levels. Results: Maximum callus induction 100, 87.5, and 85.33% resulted in the medium supplemented with 30 g/l sucrose, pH maintained at 5.5, and inoculated with 1.25g inoculum, respectively. Similarly, the highest fresh weights 65.00, 75.50, and 50.53 g/l were noted in a medium fortified with 40 g/l sucrose, inoculated 1.25g inoculum, and 6.0 pH level, respectively. However, the callus developed in a medium containing 50 g/l sucrose was found to be highly antibacterial potent with 27.3 and 26.5 mm inhibition zone against P. vulgaris and B. subtilis, respectively. Similarly, the callus grown on a medium inoculated with 1.00 g inoculum resulted in maximum antibacterial potential against S. aureus and P. vulgaris with 25 and 23.72 mm inhibition zone, respectively. However, in the case of pH levels, the medium maintained at 6.5 pH showed maximum antibacterial activity against P. vulgaris, B.subtilis, and E.coli with 27.9, 25, and 23.72 mm, respectively. The ethyl acetate extract of Stevia callus and leaves did not show antibacterial potential against Xanthomonas campestris and Clavebactor michiganensis. In the entire experiment, the standard antibacterial agent Streptomycin showed the highest inhibition zones among the rest of the callus extract; however, the pure dimethyl sulfoxide (DMSO) caused no inhibitory zone against any bacteria. Conclusion: From these findings, it is concluded that among various levels, sucrose @ 40 g L⁻¹, pH 6.0, and inoculums at 0.75 g were found best for most of the growth and quality attributes, including fresh weight, dry weight, and antibacterial activities and therefore can be recommended for callus proliferation and antibacterial potential of Stevia rebaudiana.

Keywords: Stevia rebaudiana, Steviol Glycosides, callus, Xanthomonas campestris

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Authors: Isabel Lopez-Oliva, Akshay Paropkari, Shweta Saraswat, Stefan Serban, Paola de Pablo, Karim Raza, Andrew Filer, Iain Chapple, Thomas Dietrich, Melissa Grant, Purnima Kumar

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Objective: We aimed to explicate the role of the subgingival microbiome in the causal link between rheumatoid arthritis (RA) and periodontitis (PD). Methods: Subjects with/without RA and with/without PD were randomized for treatment with scaling and root planing (SRP) or oral hygiene instructions. Subgingival biofilm, gingival crevicular fluid, and serum were collected at baseline and at 3- and 6-months post-operatively. Correlations were generated between 72 million 16S rDNA sequences, immuno-inflammatory mediators, circulating antibodies to oral microbial antigens, serum inflammatory molecules, and clinical metrics of RA. The dynamics of inter-microbial and host-microbial interactions were modeled using differential network analysis. Results: RA superseded periodontitis as a determinant of microbial composition, and DAS28 score superseded the severity of periodontitis as a driver of microbial assemblages (p=0.001, ANOSIM). RA subjects evidenced higher serum anti-PPAD (p=0.0013), anti-Pg-enolase (p=0.0031), anti-RPP3, anti- Pg-OMP and anti- Pi-OMP (p=0.001) antibodies than non-RA controls (with and without periodontitis). Following SRP, bacterial networks anchored by IL-1b, IL-4, IL-6, IL-10, IL-13, MIP-1b, and PDGF-b underwent ≥5-fold higher rewiring; and serum antibodies to microbial antigens decreased significantly. Conclusions: Our data suggest a circular relationship between RA and PD, beginning with an RA-influenced dysbiosis within the healthy subgingival microbiome that leads to exaggerated local inflammation in periodontitis and circulating antibodies to periodontal pathogens and positive correlation between severity of periodontitis and RA activity. Periodontal therapy restores host-microbial homeostasis, reduces local inflammation, and decreases circulating microbial antigens. Our data highlights the importance of integrating periodontal care into the management of RA patients.

Keywords: rheumatoid arthritis, periodontal, subgingival, DNA sequence analysis, oral microbiome

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Authors: Belinda Vega, Claudio Alvarez, Héctor Flores, Marcia Oliva, Katherine Alveal, Teresa Toro, María José Tapia, Fanny Guzmán

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With the increase in global temperatures and the decrease of oxygen (O2) concentration in the oceans, fish cultures are exposed to frequent fluctuations in dissolved O2 (DO) concentration that can cause chronic stress in the animals, altering the normal functioning of their immune system and making them vulnerable to infections, consequently increasing morbidity and mortality in the farms with economic losses. The mucosal organs (skin -and mucus-, gills, gut, and nasal mucosa) are the first line of defense of the fish against pathogens. Therefore, the objective of this study is to evaluate the effect of hypoxia on the antimicrobial activity of epidermal mucus from corvina drum (Cilus Gilberti), a native marine species with the potential for the diversification of aquaculture in Chile. To achieve this, the epidermal mucus of juveniles (~220g) kept under normoxia (7 mg/L DO) and hypoxia (2 mg/L DO) environmental conditions was collected after 6 weeks, as well as after 6 days of intraperitoneal inoculation with lipopolysaccharide from Vibrio anguillarum to induce an immune response in the fish. Total protein extracts of the mucus were used for bactericidal activity and lysozyme and peroxidase activity assays. Although the mucus from both experimental groups showed inhibitory effects on the bacterial growth of Vibrio anguillarum and Vibrio ordalli, this effect was more long-lasting in the normoxia group. We also observed a notable reduction in the presence of lysozyme in the mucus from fish exposed to hypoxia, with no differences in peroxidase content. Future proteomic studies of corvina mucus associated with the environmental conditions studied in this work will allow the isolation and identification of peptides with antimicrobial activity, those responsible for the results obtained. This will help establish strategies aimed at minimizing the impacts of hypoxia on the defense responses of corvina drum against potential pathogens. Funding: FONDECYT 3200440 and FONDECYT 1210056

Keywords: Cilus gilberti, mucus, antimicrobial activity, HYPOXIA

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Authors: Graziella Ziino, Filippo Giarratana, Stefania Maria Marotta, Alessandro Giuffrida, Antonio Panebianco

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In Europe, North America and Japan, campylobacteriosis is one of the leading food-borne bacterial illnesses, often related to the consumption of poultry meats and/or by-products. The aim of this study was the evaluation of Campylobacter contamination of poultry meats marketed in Sicily (Italy) using both traditional methods and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS). MALDI-TOF MS is considered a promising rapid (less than 1 hour) identification method for food borne pathogens bacteria. One hundred chicken and turkey meat preparations (no. 68 hamburgers, no. 21 raw sausages, no. 4 meatballs and no. 7 meat rolls) were taken from different butcher’s shops and large scale retailers and submitted to detection/enumeration of Campylobacter spp. according to EN ISO 10272-1:2006 and EN ISO 10272-2:2006. Campylobacter spp. was detected with general low counts in 44 samples (44%), of which 30 from large scale retailers and 14 from butcher’s shops. Chicken meats were significantly more contaminated than turkey meats. Among the preparations, Campylobacter spp. was found in 85.71% of meat rolls, 50% of meatballs, 44.12% of hamburgers and 28.57% of raw sausages. A total of 100 strains, 2-3 from each positive samples, were isolated for the identification by phenotypic, biomolecular and MALDI-TOF MS methods. C. jejuni was the predominant strains (63%), followed by C. coli (33%) and C. lari (4%). MALDI-TOF MS correctly identified 98% of the strains at the species level, only 1% of the tested strains were not identified. In the last 1%, a mixture of two different species was mixed in the same sample and MALDI-TOF MS correctly identified at least one of the strains. Considering the importance of rapid identification of pathogens in the food matrix, this method is highly recommended for the identification of suspected colonies of Campylobacteria.

Keywords: campylobacter spp., Food Microbiology, matrix-assisted laser desorption ionization-time of flight mass spectrometry, rapid microbial identification

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Authors: Evelyn Osehontue Uroro, Richard Bright, Jing Yang Quek, Krasimir Vasilev

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Bacterial infections have been a challenge both in the public and private sectors. The colonization of bacteria often occurs in medical devices such as catheters, heart valves, respirators, and orthopaedic implants. When biomedical devices are inserted into patients, the deposition of macromolecules such as fibrinogen and immunoglobin on their surfaces makes it easier for them to be prone to bacteria colonization leading to the formation of biofilms. The formation of biofilms on medical devices has led to a series of device-related infections which are usually difficult to eradicate and sometimes cause the death of patients. These infections require surgical replacements along with prolonged antibiotic therapy, which would incur additional health costs. It is, therefore, necessary to prevent device-related infections by inhibiting the formation of biofilms using intelligent technology. Antibiotic resistance of bacteria is also a major threat due to overuse. Different antimicrobial agents have been applied to microbial infections. They include conventional antibiotics like rifampicin. The use of conventional antibiotics like rifampicin has raised concerns as some have been found to have hepatic and nephrotoxic effects due to overuse. Hence, there is also a need for proper delivery of these antibiotics. Different techniques have been developed to encapsulate and slowly release antimicrobial agents, thus reducing host cytotoxicity. Examples of delivery systems are solid lipid nanoparticles, hydrogels, micelles, and polymeric nanoparticles. The different ways by which drugs are released from polymeric nanoparticles include diffusion-based release, elution-based release, and chemical/stimuli-responsive release. Polymeric nanoparticles have gained a lot of research interest as they are basically made from biodegradable polymers. An example of such a biodegradable polymer is polycaprolactone (PCL). PCL degrades slowly by hydrolysis but is often sensitive and responsive to stimuli like enzymes to release encapsulants for antimicrobial therapy. This study presents the synthesis of PCL nanoparticles loaded with rifampicin and the on-demand release of rifampicin for treating staphylococcus aureus infections.

Keywords: enzyme, Staphylococcus aureus, PCL, rifampicin

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Authors: Berkas Khaoula, Chaoui Kamel

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Polyethylene (PE) pipes are one of the best options for water and gas transmission networks. The main reason for such a choice is its high-quality performance in service conditions over long periods of time. PE pipes are installed in contact with different soils having various chemical compositions with confirmed aggressiveness. As a result, PE pipe surfaces undergo unwanted oxidation reactions. Usually, the polymer mixture is designed to include some additives, such as anti-oxidants, to inhibit or reduce the degradation effects. Some other additives are intended to increase resistance to the ESC phenomenon associated with polymers (ESC: Environmental Stress Cracking). This situation occurs in contact with aggressive external environments following different contaminations of soil, groundwater and transported fluids. In addition, bacterial activity and other physical or chemical media, such as temperature and humidity, can play an enhancing role. These conditions contribute to modifying the PE pipe structure and degrade its properties during exposure. In this work, the effect of distilled water, sodium hypochlorite (bleach), diluted sulfuric acid (H2SO4) and toluene-methanol (TM) mixture are studied when extruded PE samples are exposed to those environments for given periods. The chosen exposure periods are 7, 14 and 28 days at room temperature and in sealed glass containers. Post-exposure observations and ISO impact tests are presented as a function of time and chemical medium. Water effects are observed to be limited in explaining such use in real applications, whereas the changes in TM and acidic media are very significant. For the TM medium, the polymer toughness increased drastically (from 15.95 kJ/m² up to 32.01 kJ/m²), while sulfuric acid showed a steady augmentation over time. This situation may correspond to a hardening phenomenon of PE increasing its brittleness and its ability for structural degradation because of localized oxidation reactions and changes in crystallinity.

Keywords: polyethylene, toluene-methanol mixture, environmental stress cracking, degradation, impact resistance

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Authors: S. V. Ukwungwu, A. J. Abbas, G. G. Nasr

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The increasing high price of natural gas and oil with attendant increase in energy demand on world markets in recent years has stimulated interest in recovering residual oil saturation across the globe. In order to meet the energy security, efforts have been made in developing new technologies of enhancing the recovery of oil and gas, utilizing techniques like CO2 flooding, water injection, hydraulic fracturing, surfactant flooding etc. Surfactant flooding however optimizes production but poses risk to the environment due to their toxic nature. Amongst proven records that have utilized other type of bacterial in producing biosurfactants for enhancing oil recovery, this research uses a technique to combine biosurfactants that will achieve a scale of EOR through lowering interfacial tension/contact angle. In this study, three biosurfactants were produced from three Bacillus species from freeze dried cultures using sucrose 3 % (w/v) as their carbon source. Two of these produced biosurfactants were screened with the TEMCO Pendant Drop Image Analysis for reduction in IFT and contact angle. Interfacial tension was greatly reduced from 56.95 mN.m-1 to 1.41 mN.m-1 when biosurfactants in cell-free culture (Bacillus licheniformis) were used compared to 4. 83mN.m-1 cell-free culture of Bacillus subtilis. As a result, cell-free culture of (Bacillus licheniformis) changes the wettability of the biosurfactant treatment for contact angle measurement to more water-wet as the angle decreased from 130.75o to 65.17o. The influence of microbial treatment on crushed rock samples was also observed by qualitative wettability experiments. Treated samples with biosurfactants remained in the aqueous phase, indicating a water-wet system. These results could prove that biosurfactants can effectively change the chemistry of the wetting conditions against diverse surfaces, providing a desirable condition for efficient oil transport in this way serving as a mechanism for EOR. The environmental friendly effect of biosurfactants applications for industrial purposes play important advantages over chemically synthesized surfactants, with various possible structures, low toxicity, eco-friendly and biodegradability.

Keywords: bacillus, biosurfactant, enhanced oil recovery, residual oil, wettability

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Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi

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Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

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Authors: Godfrey Muiya Mukala

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This research looks at an array of fields in Sub-Saharan Africa comprising agriculture, food enterprises, medicine, organisms genetically modified, microbiology, and nanotechnology that can be gained from biotechnological research and development. Findings into dangerous organisms, mainly bacterial germs, rickettsia, fungi, parasites, or organisms that are genetically engineered, have immensely posed questions attributed to the biological danger they bring forth to human beings and the environment because of their uncertainties. In addition, the recurrence of previously managed diseases or the inception of new diseases are connected to biosafety challenges, especially in rural set-ups in low and middle-income countries. Notably, biotechnology laboratories are required to adopt biosafety measures to protect their workforce, community, environment, and ecosystem from unforeseen materials and organisms. Sensitization and inclusion of educational frameworks for laboratory workers are essential to acquiring a solid knowledge of harmful biological agents. This is in addition to human pathogenicity, susceptibility, and epidemiology to the biological data used in research and development. This article reviews and analyzes research intending to identify the proper implementation of universally accepted practices in laboratory safety and biological hazards. This research identifies ideal microbiological methods, adequate containment equipment, sufficient resources, safety barriers, specific training, and education of the laboratory workforce to decrease and contain biological hazards. Subsequently, knowledge of standardized microbiological techniques and processes, in addition to the employment of containment facilities, protective barriers, and equipment, is far-reaching in preventing occupational infections. Similarly, reduction of risks and prevention may be attained by training, education, and research on biohazards, pathogenicity, and epidemiology of the relevant microorganisms. In this technique, medical professionals in rural setups may adopt the knowledge acquired from the past to project possible concerns in the future.

Keywords: sub-saharan africa, biotechnology, laboratory, infections, health

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Authors: Zuliani Mahmood, Thirumulu Ponnuraj Kannan, Yean Yean Chan, Salahddin A. Al-Hudhairy

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Caries is the most common childhood disease which develops due to disturbances in the physiological equilibrium in the dental plaque resulting in demineralization of tooth structures. Plaque and dentine samples were collected from three different tooth surfaces representing caries progression (intact, over carious lesion and dentine) in children with early childhood caries (ECC, n=36). In caries free (CF) children, plaque samples were collected from sound tooth surfaces at baseline and after one year (n=12). The genomic DNA was extracted from all samples and subjected to 16S rRNA PCR amplification. The end products were cloned into pCR®2.1-TOPO® Vector. Five randomly selected positive clones collected from each surface were sent for sequencing. Identification of the bacterial clones was performed using BLAST against GenBank database. In the ECC group, the frequency of Lactobacillus sp. detected was significantly higher in the dentine surface (p = 0.031) than over the cavitated lesion. The highest frequency of bacteria detected in the intact surfaces was Fusobacterium nucleatum subsp. polymorphum (33.3%) while Streptococcus mutans was detected over the carious lesions and dentine surfaces at a frequency of 33.3% and 52.7% respectively. Fusobacterium nucleatum subsp. polymorphum was also found to be highest in the CF group (41.6%). Follow up at the end of one year showed that the frequency of Corynebacterium matruchotii detected was highest in those who remained caries free (16.6%), while Porphyromonas catoniae was highest in those who developed caries (25%). In conclusion, Streptococcus mutans and Porphyromonas catoniae are strongly associated with caries progression, while Lactobacillus sp. is restricted to deep carious lesions. Fusobacterium nucleatum subsp. polymorphum and Corynebacterium matruchotii may play a role in sustaining the healthy equilibrium in the dental plaque. These identified bacteria show promise as potential biomarkers in diagnosis which could help in the management of dental caries in children.

Keywords: early childhood caries, genotypic identification, oral bacteria, 16S rRNA

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Authors: Wanbao Chen, Qianqian Yao, Zhenming Zhou

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Whole grains with intact pericarp are largely resistant to digestion by ruminants because entire kernels are not conducive to bacterial attachment. But processing methods makes the starch more accessible to microbes, and increases the rate and extent of starch degradation in the rumen. To estimate the feasibility of applying a steam-flaking as the processing technique of grains for ruminants, cereal grains (maize, wheat, barley and sorghum) were processed by steam-flaking (steam temperature 105°C, heating time, 45 min). And chemical analysis, in vitro gas production, volatile fatty acid concentrations, and energetic values were adopted to evaluate the effects of steam-flaking. In vitro cultivation was conducted for 48h with the rumen fluid collected from steers fed a total mixed ration consisted of 40% hay and 60% concentrates. The results showed that steam-flaking processing had a significant effect on the contents of neutral detergent fiber and acid detergent fiber (P < 0.01). The concentration of starch gelatinization degree in all grains was also great improved in steam-flaking grains, as steam-flaking processing disintegrates the crystal structure of cereal starch, which may subsequently facilitate absorption of moisture and swelling. Theoretical maximum gas production after steam-flaking processing showed no great difference. However, compared with intact grains, total gas production at 48 h and the rate of gas production were significantly (P < 0.01) increased in all types of grain. Furthermore, there was no effect of steam-flaking processing on total volatile fatty acid, but a decrease in the ratio between acetate and propionate was observed in the current in vitro fermentation. The present study also found that steam-flaking processing increased (P < 0.05) organic matter digestibility and energy concentration of the grains. The collective findings of the present study suggest that steam-flaking processing of grains could improve their rumen fermentation and energy utilization by ruminants. In conclusion, the utilization of steam-flaking would be practical to improve the quality of common cereal grains.

Keywords: cereal grains, gas production, in vitro rumen fermentation, steam-flaking processing

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Authors: Qunying Yuan, Manjula Bomma, Adrian Rhoden, Zhigang Xiao

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Selenium is an essential micronutrient for all mammals and plays an important role in maintaining human physiological functions. The potential applications of selenium as food supplements, cancer-prevention, antimicrobial and anti-inflammatory agents have been investigated in biomedicine and food sciences. Nanoscale of selenium is of particular interest due to its better biocompatibility, higher bioavailability, lower toxicity, more homogeneous distribution, and presumptive controlled release of substances. The objective of this study is to explore whether selenium nanoparticle (SeNP) has the potential to be used as a food preservative to reduce food spoilage. SeNPs were synthesized through ascorbic acid reduction of sodium selenite using the bovine serum albumin (BSA) as capping and stabilizing agent. The chemically synthesized SeNPs had a spherical conformation and a size of 22.8 ± 4.7 nm. FTIR analysis confirmed that the nanoparticles were covered with BSA. We further tested the antimicrobial activity of these SeNPs against common food-borne bacteria. Colony forming unit assay showed that SeNPs exhibited good inhibition on the growth of Listeria Monocytogens (ATCC15313), Staphylococcus epidermidis (ATCC 700583) starting at 0.5µg/mL, but only a moderate inhibitory effect on the growth of Staphylococcus aureus (ATCC12600) and Vibrio alginolyticus (ATCC 33787) at a concentration higher than 10µg/mL and 2.5µg/mL, respectively. There was a mild effect against the growth Salmonella enterica (ATCC19585) when the concentration reached 15µg/mL. No inhibition was observed in the growth of Enterococcus faecalis (ATCC 19433). Surprisingly, SeNPs appeared to promote the growth of Vibrio parahaemolyticus (ATCC43996) and Salmonella enterica (ATCC49284) at 30 µg/mL and above. Our preliminary data suggested that the chemically synthesized SeNPs may be able to inhibit some food-borne bacteria, and SeNP as a food preservative should be used with caution. We will explore the mechanisms of the inhibitory action of chemically synthesized SeNPs on bacterial growth and whether the SeNPs are able to inhibit the development of biofilm and antibiotic resistance.

Keywords: antimicrobial, food-borne bacteria, nanoparticles, selenium

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Authors: Stelvin Sebastian, Andriya Annie Tom, Joyalanna Babu, Merin Joshy

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Introduction: The worldwide increase in the use of antibiotics in poultry and livestock industry to treat and prevent bacterial diseases and as growth promoters in feeds has led to the problem of development of antibiotic resistance both in animals and human population. Aim: To study the pattern of antibiotic use and prevalence of multidrug-resistant Escherichia coli in poultry chickens in selected farms in Muvattupuzha and to compare the spread of multidrug-resistant bacteria from poultry environment to UTI patients. Methodology: Two farms from each of 6 localities in Muvattupuzha were selected. A questionnaire on the pattern of antibiotic use and various farming practices were surveyed from farms. From each farm, 60samples of fresh fecal matter, litter from inside, litter from the outside shed, agricultural soil and control soil were collected, and antimicrobial susceptibility testing of E. coli was done. Antibiogram of UTI patients was collected from the secondary care hospital included in the study, and those were compared with resistance patterns of poultry samples. Results: From survey response antibiotics such as ofloxacin, enrofloxacin, levofloxacin, ciprofloxacin, colistin, ceftriaxone, neomycin, cephalexin, and oxytetracycline were used for treatment and prevention of infections in poultry. 31of 48 samples (51.66%) showed E. coli growth. 7 of 15 antibiotics (46.6%) showed resistance. Ampicillin, amoxicillin, meropenem, tetracycline showed 100% resistance to all samples. Statistical analysis confirmed similar resistance pattern in the poultry environment and UTI patients for antibiotics such as ampicillin, amoxicillin, amikacin, and ofloxacin. Conclusion: E. coli were resistant not only to extended-spectrum beta-lactams but also to carbapenems, which may be disseminated to the environment where litter was used as manure. This may due to irrational use of antibiotics in chicken or from their use in poultry feed as growth promoters. The study concludes the presence of multidrug-resistant E.coli in poultry and its spread to environment and humans, which may cause potentially serious implications for human health.

Keywords: multidrug resistance, escherichia coli, urinary tract infection, poultry

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Authors: Anna-Mari Kok, Carel B. Oosthuizen, Namrita Lall

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Tuberculosis (TB) is a disease caused by the bacterial pathogen Mycobacterium tuberculosis. It is associated with high mortality rates in both developing and developed countries. Many higher plants are found that are medicinally associated with tuberculosis infection. Plants belonging to thirteen families were selected, based on their traditional usage for tuberculosis and its associated symptoms. Eight plants showed the best antimycobacterial activities (MIC-value ≤ 500.0 µg/ml) against M. tuberculosis H37Rv. LS was found to have a minimum inhibitory concentration (MIC) of 125 µg/ml whereas, Tulbaghia violacea, Heteromorpha arborescens, Sutherlandia frutescens, Eucalyptus deglupta, and Plectranthus neochilus were found to have a MIC value of 250 µg/ml against M. tuberculosis H37Rv. Cytotoxicity values on U937 and HepG2 cells were obtained and the IC50 values ranged between 40 ±4.30 and > 400 µg/ml for the U937 cell line and 72.4 ±1.50 and > 400 µg/ml for the HepG2 cell line. Heteromorpha arborescens had the lowest IC50 value in both cell lines and therefore showed moderate levels of toxicity. Of the 19 samples that underwent the 2, 2- diphenyl- 1- picrylhydrazyl (DPPH) antioxidant assay, Eucalyptus deglupta and Melianthus major showed significant free radical scavenging activities with concentrations of 1.33 and 1.32 µg/ml respectively for the inhibition of DPPH. Hepatotoxicity induced by acetaminophen identified Searsia lancea with hepatoprotective activity of 59.37% at a ¼ IC50 concentration. Out of the 7 samples that were investigated for their immunomodulatory capabilities, Eucalyptus deglupta produced the most IL-12 with Sutherlandia frutescens also showing positive results for IL-12 production. In the present study, Eucalyptus deglupta showed the most promising results with good activity against M. tuberculosis with an MIC-value of 250 µg/ml. It also has potent antioxidant activity with an IC50 value of 1.33 µg/ml. This sample also stimulated high production of the cytokine, IL-12. Searsia lancea showed moderate antimycobacterial acticvity with an MIC-value of 500 µg/ml. The antioxidant potential also showed promising results with an IC50 value of 4.50 µg/ml. The hepatoprotective capability of Searsia lancea was 59.34% at a ¼ IC50 concentration. Another sample Sutherlandia frutescens showed effective antimycobacterial activity with an MIC-value of 250 µg/ml. It also stimulated production of IL-12 with 13.43 pg/ml produced. These three samples can be considered for further studies for the consideration as adjuvants for current tuberculosis treatment.

Keywords: adjuvant, hepatoprotection, immunomodulation, tuberculosis

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Authors: Rachel Fanelwa Ajayi, Anovuyo Jonnas, Emmanuel I. Iwuoha

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Tuberculosis (TB) is an infectious disease caused by the bacterium (Mycobacterium tuberculosis) which has a predilection for lung tissue due to its rich oxygen supply. The mycobacterial cell has a unique innate characteristic which allows it to resist human immune systems and drug treatments; hence, it is one of the most difficult of all bacterial infections to treat, let alone to cure. At the same time, multi-drug resistance TB (MDR-TB) caused by poorly managed TB treatment, is a growing problem and requires the administration of expensive and less effective second line drugs which take much longer treatment duration than fist line drugs. Therefore, to acknowledge the issues of patients falling ill as a result of inappropriate dosing of treatment and inadequate treatment administration, a device with a fast response time coupled with enhanced performance and increased sensitivity is essential. This study involved the synthesis of electroactive platforms for application in the development of nano-biosensors suitable for the appropriate dosing of clinically diagnosed patients by promptly quantifying the levels of the TB drug; Isonaizid. These nano-biosensors systems were developed on gold surfaces using the enzyme N-acetyletransferase 2 coupled to the cysteamine modified poly(8-anilino-1-napthalene sulphonic acid)/zinc oxide nanocomposites. The morphology of ZnO nanoparticles, PANSA/ZnO nano-composite and nano-biosensors platforms were characterized using High-Resolution Transmission Electron Microscopy (HRTEM) and High-Resolution Scanning Electron Microscopy (HRSEM). On the other hand, the elemental composition of the developed nanocomposites and nano-biosensors were studied using Fourier Transform Infra-Red Spectroscopy (FTIR) and Energy Dispersive X-Ray (EDX). The electrochemical studies showed an increase in electron conductivity for the PANSA/ZnO nanocomposite which was an indication that it was suitable as a platform towards biosensor development.

Keywords: N-acetyletransferase 2, isonaizid, tuberculosis, zinc oxide

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Authors: Marina Arino Martin, John McEvoy, Eakalak Khan

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Endoxifen is an active metabolite responsible for the effectiveness of tamoxifen, a chemotherapeutic drug widely used for endocrine responsive breast cancer and chemo-preventive long-term treatment. Tamoxifen and endoxifen are not completely metabolized in human body and are actively excreted. As a result, they are released to the water environment via wastewater treatment plants (WWTPs). The presence of tamoxifen in the environment produces negative effects on aquatic lives due to its antiestrogenic activity. Because endoxifen is 30-100 times more potent than tamoxifen itself and also presents antiestrogenic activity, its presence in the water environment could result in even more toxic effects on aquatic lives compared to tamoxifen. Data on actual concentrations of endoxifen in the environment is limited due to recent discovery of endoxifen pharmaceutical activity. However, endoxifen has been detected in hospital and municipal wastewater effluents. The detection of endoxifen in wastewater effluents questions the treatment efficiency of WWTPs. Studies reporting information about endoxifen removal in WWTPs are also scarce. There was a study that used chlorination to eliminate endoxifen in wastewater. However, an inefficient degradation of endoxifen by chlorination and the production of hazardous disinfection by-products were observed. Therefore, there is a need to remove endoxifen from wastewater prior to chlorination in order to reduce the potential release of endoxifen into the environment and its possible effects. The aim of this research is to isolate and identify bacteria strain(s) capable of degrading endoxifen into less hazardous compound(s). For this purpose, bacteria strains from WWTPs were exposed to endoxifen as a sole carbon and nitrogen source for 40 days. Bacteria presenting positive growth were isolated and tested for endoxifen biodegradation. Endoxifen concentration and by-product formation were monitored. The Monod kinetic model was used to determine endoxifen biodegradation rate. Preliminary results of the study suggest that isolated bacteria from WWTPs are able to growth in presence of endoxifen as a sole carbon and nitrogen source. Ongoing work includes identification of these bacteria strains and by-product(s) of endoxifen biodegradation.

Keywords: biodegradation, bacterial degraders, endoxifen, wastewater

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Authors: Apichaya Aneksampant, Xuefei Tu, Masami Fukushima, Mitsuo Yamamoto

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The restoration of seaweed beds recovery has been developed using a fertilization technique for supplying dissolved iron to barren coastal areas. The fertilizer is composed of iron oxides as a source of iron and compost as humic substance (HS) source, which can serve as chelator of iron to stabilize the dissolved species under oxic seawater condition. However, elution mechanisms of iron from iron oxide surfaces have not sufficiently elucidated. In particular, roles of microbial activities in the elution of iron from the fertilizer are not sufficiently understood. In the present study, a fertilizer (iron oxide/compost = 1/1, v/v) was incubated in a water tank at Mashike coast, Hokkaido Japan. Microorganisms in the 6-month fertilizer were isolated and identified as Exiguobacterium oxidotolerans sp. (T-2-2). The identified bacteria were inoculated to perform iron elution test in a postgate B medium, prepared in artificial seawater. Hematite was used as a model iron oxide and anthraquinone-2,7-disolfonate (AQDS) as a model for HSs. The elution test performed in presence and absence of bacteria inoculation. ICP-AES was used to analyze total iron and a colorimetric technique using ferrozine employed for the determination of ferrous ion. During the incubation period, sample contained hematite and T-2-2 in both presence and absence of AQDS continuously showed the iron elution and reached at the highest concentration after 9 days of incubation and then slightly decrease to stabilize within 20 days. Comparison to the sample without T-2-2, trace amount of iron was observed, suggesting that iron elution to seawater can be attributed to bacterial activities. The levels of total organic carbon (TOC) in the culture solution with hematite decreased. This may be to the adsorption of organic compound, AQDS, to hematite surfaces. The decrease in UV-vis absorption of AQDS in the culture solution also support the results of TOC that AQDS was adsorbed to hematite surfaces. AQDS can enhance the iron elution, while the adsorption of organic matter suppresses the iron elution from hematite.

Keywords: anthraquinone-2, 7-disolfonate, barren ground, E.oxidotolerans sp., hematite, humic substances, iron elution

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Authors: Mpho Mokoatle, Darlington Mapiye, James Mashiyane, Stephanie Muller, Gciniwe Dlamini

Abstract:

Great advances in high-throughput sequencing technologies have resulted in availability of huge amounts of sequencing data in public and private repositories, enabling a holistic understanding of complex biological phenomena. Sequence data are used for a wide range of applications such as gene annotations, expression studies, personalized treatment and precision medicine. However, this rapid growth in sequence data poses a great challenge which calls for novel data processing and analytic methods, as well as huge computing resources. In this work, a machine and statistical learning approach for DNA sequence classification based on $k$-mer representation of sequence data is proposed. The approach is tested using whole genome sequences of Mycobacterium tuberculosis (MTB) isolates to (i) reduce the size of genomic sequence data, (ii) identify an optimum size of k-mers and utilize it to build classification models, (iii) predict the phenotype from whole genome sequence data of a given bacterial isolate, and (iv) demonstrate computing challenges associated with the analysis of whole genome sequence data in producing interpretable and explainable insights. The classification models were trained on 104 whole genome sequences of MTB isoloates. Cluster analysis showed that k-mers maybe used to discriminate phenotypes and the discrimination becomes more concise as the size of k-mers increase. The best performing classification model had a k-mer size of 10 (longest k-mer) an accuracy, recall, precision, specificity, and Matthews Correlation coeffient of 72.0%, 80.5%, 80.5%, 63.6%, and 0.4 respectively. This study provides a comprehensive approach for resampling whole genome sequencing data, objectively selecting a k-mer size, and performing classification for phenotype prediction. The analysis also highlights the importance of increasing the k-mer size to produce more biological explainable results, which brings to the fore the interplay that exists amongst accuracy, computing resources and explainability of classification results. However, the analysis provides a new way to elucidate genetic information from genomic data, and identify phenotype relationships which are important especially in explaining complex biological mechanisms.

Keywords: AWD-LSTM, bootstrapping, k-mers, next generation sequencing

Procedia PDF Downloads 136

Authors: Darlington Mapiye, Mpho Mokoatle, James Mashiyane, Stephanie Muller, Gciniwe Dlamini

Abstract:

Great advances in high-throughput sequencing technologies have resulted in availability of huge amounts of sequencing data in public and private repositories, enabling a holistic understanding of complex biological phenomena. Sequence data are used for a wide range of applications such as gene annotations, expression studies, personalized treatment and precision medicine. However, this rapid growth in sequence data poses a great challenge which calls for novel data processing and analytic methods, as well as huge computing resources. In this work, a machine and statistical learning approach for DNA sequence classification based on k-mer representation of sequence data is proposed. The approach is tested using whole genome sequences of Mycobacterium tuberculosis (MTB) isolates to (i) reduce the size of genomic sequence data, (ii) identify an optimum size of k-mers and utilize it to build classification models, (iii) predict the phenotype from whole genome sequence data of a given bacterial isolate, and (iv) demonstrate computing challenges associated with the analysis of whole genome sequence data in producing interpretable and explainable insights. The classification models were trained on 104 whole genome sequences of MTB isoloates. Cluster analysis showed that k-mers maybe used to discriminate phenotypes and the discrimination becomes more concise as the size of k-mers increase. The best performing classification model had a k-mer size of 10 (longest k-mer) an accuracy, recall, precision, specificity, and Matthews Correlation coeffient of 72.0 %, 80.5 %, 80.5 %, 63.6 %, and 0.4 respectively. This study provides a comprehensive approach for resampling whole genome sequencing data, objectively selecting a k-mer size, and performing classification for phenotype prediction. The analysis also highlights the importance of increasing the k-mer size to produce more biological explainable results, which brings to the fore the interplay that exists amongst accuracy, computing resources and explainability of classification results. However, the analysis provides a new way to elucidate genetic information from genomic data, and identify phenotype relationships which are important especially in explaining complex biological mechanisms

Keywords: AWD-LSTM, bootstrapping, k-mers, next generation sequencing

Procedia PDF Downloads 126

Authors: Adepeju Simon-Oke, Olatunji Odeyemi, Mobolanle Oniya

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Urinary tract infection has become the most common bacterial infections in humans, both at the community and hospital settings; it has been reported in all age groups and in both sexes. This study was carried out in order to determine and evaluate the prevalence, current drug susceptibility pattern of the isolated organisms and identify the associated risk factors of UTIs among the pregnant women in Akure, Ondo State, Nigeria. A cross-sectional study was conducted on the urine of pregnant women, and socio-demographic information of the women was collected. A total of 300 clean midstream urine samples were collected, and a general urine microscopic examination and culture were carried out, the Microbact identification system was used to identify gram-negative bacteria. Out of the 300 urine samples cultured, 183(61.0%) yielded significant growth of urinary pathogens while 117(39.0%) yielded either insignificant growth or no growth of any urinary pathogen. Prevalence of UTI was significantly associated with the type of toilet used, symptoms of UTI, and previous history of urinary tract infection (p<0.05). Escherichia coli 58(31.7%) was the dominant pathogen isolated, and the least isolated uropathogens were Citrobacter freudii and Providencia retgerri 2(1.1%) respectively. Gram-negative bacteria showed 77.6%, 67.9%, and 61.2% susceptibility to ciprofloxacin, augmentin, and chloramphenicol, respectively. Resistance against septrin, chloramphenicol, sparfloxacin, amoxicillin, augmentin, gentamycin, pefloxacin, trivid, and streptomycin was observed in the range of 23.1 to 70.1%. Gram-positive uropathogens isolated showed high resistance to amoxicillin (68.4%) and high susceptibility to the remaining nine antibiotics in the range 65.8% to 89.5%. This study justifies that pregnant women are at high risk of UTI. Therefore screening of pregnant women during antenatal clinics should be considered very important to avoid complications. Health education with regular antenatal and personal hygiene is recommended as precautionary measures to UTI.

Keywords: pregnant women, prevalence, risk factor, UTIs

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Authors: Islam Nowisser, Noha Farag, Mohamed El Azizi

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Helicobacter pylori is a highly important human pathogen which inhabits about 50% of the population worldwide. Infection with this bacteria is very hard to treat, with high probability of recurrence. H. pylori causes severe gastric diseases, including peptic ulcer, gastritis, and gastric cancer, which has been linked to chronic inflammation. The infection has been reported to be associated with high levels of pro-inflammatory cytokines, especially IL-1β and TNF-α. The aim of the current study is to investigate the molecular mechanisms by which H. pylori activates NLRP3 inflammasome and its contribution to Il-1 β production in an innate cellular model. H. pylori PMSS1 and G27 standard strains, as well as the PMSS1 isogenic mutant strain PMSS1ΔVacA and G27ΔVacA, G27ΔCagA in addition to clinical isolates obtained from biopsy samples from the antrum and corpus mucosa of chronic gastritis patients, were used to establish infection in RAW-264.7 macrophages. The production levels of TNF-α and IL-1β was assessed using ELISA. Since expression of these cytokines is often regulated by the transcription factor complex, nuclear factor-kB (NF-kB), the activation of NF-κB in H. pylori infected cells was also evaluated by luciferase assay. Genomic DNA was extracted from bacterial cultures of H. pylori clinical isolates as well as the standard strains and their corresponding mutants, where they were evaluated for the cagA pathogenicity island and vacA expression. The correlation between these findings and expression of the cagA Pathogenicity Island and vacA in the bacteria was also investigated. The results showed IL-1β, and TNF-α production significantly increased in raw macrophages following H. pylori infection. The cagA+ and vacA+ H. pylori strains induced significant production of IL-1β compared to cagA- and vacA- strains. The activation pattern of NF-κB was correlated in the isolates to their cagA and vacA expression profiles. A similar finding could not be confirmed for TNF-α production. Our study shows the ability of H. pylori to activate NF-kB and induce significant IL-1β production as a possible mechanism for the augmented inflammatory response seen in subjects infected with cagA+ and vacA+ H. pylori strains that would lead to the progression to more severe form of the disease.

Keywords: Helicobacter pylori, IL-1β, inflammatory cytokines, nuclear factor KB, TNF-α

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Authors: Mohammed H. Abu-Dieyeh, Mohammad M. Zalloum

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Plant growth-promoting rhizobacteria (PGPR) are known to enhance plant growth and/or reduce plant damage due to soil-borne pathogens. Tomato is the highest consumable vegetable world-wide including Jordan. Fusarium oxysporum is a pathogen that causes well-known damages and losses to many vegetable crops including tomato. In this study, purification of 112 isolates of PGPR strains from rhizosphere environment of different regions in Jordan was accomplished. All bacterial isolates were In-vitro screened for antagonistic effects against F. oxysporum. The eleven most effective isolates that caused 30%-50% in-vitro growth reduction of F. oxysporum were selected. 8 out of 11 of these isolates were collected from Al-Halabat (arid-land). 7 isolates of Al-Halabat exerted 40-54% In-vitro growth reduction of F. oxysporum. Four-week-old seedlings of tomato cultivar (Anjara, the most susceptible indigenous cultivar to F. oxysporum) treated with PGPR5 (Bacillus amyloliquefaciens), and exposed to F. oxysporum, showed no disease symptoms and no significant changes in biomasses or chlorophyll contents indicating a non-direct mechanism of action of PGPR on tomato plants. However PGPR3 (Bacillus sp.), PGPR4 (Bacillus cereus), and PGPR38 (Paenibacillus sp.) treated plants or PGPR treated and exposed to F. oxysporum showed a significant increasing growth of shoot and root biomasses as well as chlorophyll contents of leaves compared to control untreated plants or plants exposed to the fungus without PGPR treatment. A significant increase in number of flowers per plant was also recorded in all PGPR treated plants. The characterization of rhizobacterial strains were accomplished using 16S rRNA gene sequence analysis in addition to microscopic characterization. Further research is necessary to explore the potentiality of other collected PGPR isolates on tomato plants in addition to investigate the efficacy of the identified isolates on other plant pathogens and then finding a proper and effective methods of formulation and application of the successful isolates on selected crops.

Keywords: antagonism, arid land, growth promoting, rhizobacteria, tomato

Procedia PDF Downloads 349

Authors: Yassir Adam Shuaib, Stefan Niemann, Eltahir Awad Khalil, Ulrich Schaible, Lothar Heinz Wieler, Mohammed Ahmed Bakhiet, Abbashar Osman Mohammed, Mohamed Abdelsalam Abdalla, Elvira Richter

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Tuberculosis (TB) is a chronic bacterial disease of humans and animals and it is characterized by the progressive development of specific granulomatous tubercle lesions in affected tissues. In a six-month study, from June to November 2014, a total of 2,304 carcasses of cattle, camel, sheep, and goats slaughtered at East and West Gaash slaughterhouses, Kassala, were investigated during postmortem, in parallel, 101 sputum samples from TB suspected patients at Kassala and El-Gadarif Teaching Hospitals were collected in order to investigate tuberculosis in animals and humans. Only 0.1% carcasses were found with suspected TB lesions in the liver and lung and peritoneal cavity of two sheep and no tuberculous lesions were found in the carcasses of cattle, goats or camels. All samples, tissue lesions and sputum, were decontaminated by the NALC-NaOH method and cultured for mycobacterial growth at the NRZ for Mycobacteria, Research Center Borstel, Germany. Genotyping and molecular characterization of the grown strains were done by line probe assay (GenoType CM and MTBC) and 16S rDNA, rpoB gene, and ITS sequencing, spoligotyping, MIRU-VNTR typing and next generation sequencing (NGS). Culture of the specimens revealed growth of organisms from 81.6% of all samples. Mycobacterium tuberculosis (76.2%), M. intracellulare (14.2%), mixed infection with M. tuberculosis and M. intracellulare (6.0%) and mixed infection with M. tuberculosis and M. fortuitum and with M. intracellulare and unknown species (1.2%) were detected in the sputum samples and unknown species (1.2%) were detected in the samples of one of the animals tissues. From the 69 M. tuberculosis strains, 25 (36.2%) were showing either mono-drug-resistant or multi-drug-resistant or poly-drug-resistant but none was extensively drug-resistant. In conclusion, the prevalence of TB in animals was very low while in humans M. tuberculosis-Delhi/CAS lineage was responsible for most cases and there was an evidence of MDR transmission and acquisition.

Keywords: animal, human, slaughterhouse, Sudan, tuberculosis

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Authors: Vaibhav D. Bhatt, Anju P. Kunjadia, D. S. Nauriyal, Bhumika J. Joshi, Chaitanya G. Joshi

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Metagenomic analysis of milk samples collected from local cattle breed, kankrej (Bos indicus), Gir (Bos indicus) and Crossbred (Bos indicus X Bos taurus) cattle harbouring subclinical mastitis was carried out by next-generation sequencing (NGS) 454 GS-FLX technology. Around 56 different species including members of Enterobacteriales, Pseudomonadales, Bacillales and Lactobacillales with varying abundance were detected in infected milk. The interesting presence of bacteriophages against Staphylococcus aureus, Escherichia coli, Enterobacter and Yersinia species were observed, especially Enterobacteria and E. coli phages (0∙32%) in Kankrej, Enterobacteria and Staphylococcus phages (1∙05%) in Gir and Staphylococcus phages (2∙32%) in crossbred cattle. NGS findings suggest that phages may be involved in imparting natural resistance of the cattle against pathogens. Further infected milk samples were subjected for bacterial isolation. Fourteen different isolates were identified, and DNA was extracted. Genes (Tet-K, Msr-A, and Mec-A) providing antibiotic resistance to the bacteria were screened by Polymerase Chain Reaction and results were validated with traditional antibiotic assay. Total 3 bacteriophages were isolated from nearby environment of the cattle farm. The efficacy of phages was checked against multi-drug resistant bacteria, identified by PCR. In-vivo study was carried out for phage therapy in mammary glands of female rats “Wister albino”. Mammary glands were infused with MDR isolates for 3 consecutive days. Recovery was observed in infected rats after intramammary infusion of sterile phage suspension. From day 4th onwards, level of C-reactive protein was significant increases up to day 12th . However, significant reduction was observed between days 12th to 18th post treatment. Bacteriophages have significant potential as antibacterial agents and their ability to replicate exponentially within their hosts and their specificity, make them ideal candidates for more sustainable mastitis control.

Keywords: bacteriophages, c-reactive protein, mastitis, metagenomic analysis

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Authors: Komal Vig, Syamala Soumyakrishnan, Yadav Baral

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Bypass surgery, using the autologous vein has been one of the most effective treatments for cardiovascular diseases (CVD). More recently tissue engineering including engineered vascular grafts to synthesize blood vessels is gaining usage. Dacron and ePTFE has been employed for vascular grafts, however, these does not work well for small diameter grafts (<6 mm) due to intimal hyperplasia and thrombosis. In the present study PTFE was treated with LTP to improve the endothelialization of intimal surface of graft. Scaffolds were also modified with polyvinylpyrrolidone coated silver nanoparticles (Ag-PVP) and the antimicrobial peptides, p753 and p359. Human umbilical vein endothelial cells (HUVEC) were plated on the developed scaffolds and cell proliferation was determined by the MTT assay. Cells attachment on scaffolds was visualized by microscopy. mRNA expressions levels of different cell markers were investigated using quantitative real-time PCR (qPCR). X ray photoelectron spectroscopic confirmed the introduction of oxygenated functionalities from LTP air plasma. Microscopic and MTT assays indicated increase in cell viability in LTP treated scaffolds. Gene expression studies shows enhanced expression of cell adhesion marker Integrin- α 5 gene after LTP treatment. The KB test displayed a zone of inhibition for Ag-PVP, p753 and p359 of 19mm, 14mm, and 12mm respectively. To determine toxicity of antimicrobial agents to cells, MTT Assay was performed using HEK293 cells. MTT Assay exhibited that Ag-PVP and the peptides were non-toxic to cells at 100μg/mL and 50μg/mL, respectively. Live/dead analysis and plate count of treated bacteria exhibited bacterial inhibition on develop scaffold compared to non-treated scaffold. SEM was performed to analyze the structural changes of bacteria after treatment with antimicrobial agents. Gene expression studies were conducted on RNA from bacteria treated with Ag-PVP and peptides using qRT-PCR. Based on our initial results, more scaffolds alternatives will be developed and investigated for cell growth and vascularization studies.

Keywords: low temperature plasma, vascular graft, HUVEC cells, antimicrobial

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Authors: A. Heshmati, M. Y Alikhani, M. T. Godarzi, M. R. Sadeghimanesh

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Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl₃) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Keywords: antimicrobial, antioxidant, extract, hypericum

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