Search results for: Bacteria encapsulation

Authors: Zh. Karaulova, D. Baizhigitov

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At the Aktogay deposit, the oxidized ore section has been developed since 2015; by now, the reserves of easily enriched ore are decreasing, and a large number of copper-poor, difficult-to-enrich ores has been accumulated in the dumps of the KAZ Minerals Aktogay deposit, which is unprofitable to mine using the traditional mining methods. Hence, another technology needs to be implemented, which will significantly expand the raw material base of copper production in Kazakhstan and ensure the efficient use of natural resources. Heap and dump bacterial recovery are the most acceptable technologies for processing low-grade secondary copper sulfide ores. Test objects were the copper ores of Aktogay deposit and chemolithotrophic bacteria Leptospirillum ferrooxidans (L.f.), Acidithiobacillus caldus (A.c.), Sulfobacillus Acidophilus (S.a.), which are mixed cultures were both used in bacterial oxidation systems. They can stay active in the 20-400C temperature range. These bacteria were the most extensively studied and widely used in sulfide mineral recovery technology. Biocatalytic acceleration was achieved as a result of bacteria oxidizing iron sulfides to form iron sulfate, which subsequently underwent chemical oxidation to become sulfate oxide. The following results have been achieved at the initial stage: the goal was to grow and maintain the life activity of bacterial cultures under laboratory conditions. These bacteria grew the best within the pH 1,2-1,8 range with light stirring and in an aerated environment. The optimal growth temperature was 30-33оC. The growth rate decreased by one-half for each 4-5°C fall in temperature from 30°C. At best, the number of bacteria doubled every 24 hours. Typically, the maximum concentration of cells that can be grown in ferrous solution is about 107/ml. A further step researched in this case was the adaptation of microorganisms to the environment of certain metals. This was followed by mass production of inoculum and maintenance for their further cultivation on a factory scale. This was done by adding sulfide concentrate, allowing the bacteria to convert the ferrous sulfate as indicated by the Eh (>600 mV), then diluting to double the volume and adding concentrate to achieve the same metal level. This process was repeated until the desired metal level and volumes were achieved. The final stage of bacterial recovery was the transportation and irrigation of secondary sulfide copper ores of the oxidized ore section. In conclusion, the project was implemented at the Aktogay mine since the bioleaching process was prolonged. Besides, the method of bacterial recovery might compete well with existing non-biological methods of extraction of metals from ores.

Keywords: bacterial recovery, copper ore, bioleaching, bacterial inoculum

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Authors: Pavitra Sharma, Anuradha Singh, Nupur Mathur

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There exist more than 100 trillion bacteria in the digestive system of human-beings. Such bacteria are referred to as gut microbiota. Gut microbiota comprises around 75% of our immune system. The bacteria that comprise the gut microbiota are unique to every individual and their composition keeps changing with time owing to factors such as the host’s age, diet, genes, environment, and external medication. Of these factors, the variable easiest to control is one’s diet. By modulating one’s diet, one can ensure an optimal composition of the gut microbiota yielding several health benefits. Prebiotics and probiotics are two compounds that have been considered as viable options to modulate the host’s diet. Prebiotics are basically plant products that support the growth of good bacteria in the host’s gut. Examples include garden asparagus, aloe vera etc. Probiotics are living microorganisms that exist in our intestines and play an integral role in promoting digestive health and supporting our immune system in general. Examples include yogurt, kimchi, kombucha etc. In the context of modulating the host’s diet, the key attribute of prebiotics is that they support the growth of probiotics. By developing the right combination of prebiotics and probiotics, food products or supplements can be created to enhance the host’s health. An effective combination of prebiotics and probiotics that yields health benefits to the host is referred to as synbiotics. Synbiotics comprise of an optimal proportion of prebiotics and probiotics, their application benefits the host’s health more than the application of prebiotics and probiotics used in isolation. When applied to food supplements, synbiotics preserve the beneficial probiotic bacteria during storage period and during the bacteria’s passage through the intestinal tract. When applied to the gastrointestinal tract, the composition of the synbiotics assumes paramount importance. Reason being that for synbiotics to be effective in the gastrointestinal tract, the chosen probiotic must be able to survive in the stomach’s acidic environment and manifest tolerance towards bile and pancreatic secretions. Further, not every prebiotic stimulates the growth of a particular probiotic. The prebiotic chosen should be one that not only maintains 2 balance in the host’s digestive system, but also provides the required nutrition to probiotics. Hence in each application of synbiotics, the prebiotic-probiotic combination needs to be carefully selected. Once the combination is finalized, the exact proportion of prebiotics and probiotics to be used needs to be considered. When determining this proportion, only that amount of a prebiotic should be used that activates metabolism of the required number of probiotics. It was observed that while probiotics are active is both the small and large intestine, the effect of prebiotics is observed primarily in the large intestine. Hence in the host’s small intestine, synbiotics are likely to have the maximum efficacy. In small intestine, prebiotics not only assist in the growth of probiotics, but they also enable probiotics to exhibit a higher tolerance to pH levels, oxygenation, and intestinal temperature

Keywords: microbiota, probiotics, prebiotics, synbiotics

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Authors: Michal Natan, Ori Gutman, Shlomo Margel, Ehud Banin

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Biofilm formation is a serious problem in medical and industrial settings due to the increased resistance of these communities to killing compared to free-living bacteria. This has prompted the search for agents that can inhibit both bacterial growth and biofilm formation. In this study, N-halamine rechargeable nanoparticles (NPs) were synthesized by co-polymerization of the monomer methacryl amide and the cross-linker monomer N,N-methylene bisacryl amide, and were subsequently loaded with Cl+, using bleach. The chlorinated NPs exhibited remarkable stability to organic reagents. The antibacterial mechanism of the P(MAA-MBAA)-Cl NPs involved generation of reactive oxygen species (ROS) only upon exposure to organic media, but not upon incubation in water, suggesting a specific activation. Moreover, a unique interaction of the P(MAA-MBAA)-Cl NPs with Staphylococcus aureus bacteria but not with human cells was discovered, whereby these microorganisms were all specifically targeted and marked for destruction. Finally, in collaboration with Netafim Ltd. irrigation drippers containing the P(MAA-MBAA)-Cl were incubated in the field and were shown to prevent fouling on them for 5 months as opposed to the control drippers that exhibited substantial fouling. Further, the NPs offer recharging to the surface, thus providing long-lasting protection that does not exist in the products available today. Taken together, the results demonstrate the great potential of implementing the charged NPs in devices and surfaces to prevent bacterial growth.

Keywords: bacteria, biofilm, fouling, nanoparticles

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Authors: Sayaka Ono, Ryutaro Imai, Tomoko Ehara, Tetsuya Matsumoto, Hajime Matsumura

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Background: Wound pH affects a number of important factors in wound healing. We previously measured the pH value of the exudates collected from second-degree burns and found that the increase in pH was observed in the burn wounds in which colonized by Staphylococcus spp., and the increase in pH was evident prior to the clinical findings of local infection. To investigate the relationship between the changes of pH value and bacterial growth, we performed in vitro study using Pseudomonas aeruginosa and liquid medium as a locally infected wound equivalent model. Methods: Pseudomonas aeruginosa standard strain (ATCCR 10145TM) was cultured at 37 °C environment in Luria Broth Miller medium. The absorbance rate which means the amount of bacteria was measured by a microplate reader 2300EnSpireTM). The pH was measured using pH-indicator strips (MColorpHastTM). The statistical analysis was performed using the product-moment correlation coefficient of Pearson's. Results: The absorbance rate and pH value were increased along with culture period. There was a positive correlation between pH value and absorbance rate (n = 27, Pearson's r = 0.985). Moreover, there was a positive correlation between pH value and the culture period (n = 18, Pearson's r = 0.901). The bacteria was well growth in the media from pH 6.6 to pH 8.0 and the pH of culture media converged at 8 -9 along with the bacterial growth. Conclusion: From these results, we conclude that pH value of the wound is correlated with the number of viable bacteria and bacterial growth periods.

Keywords: colonization, potential of hydrogen, Pseudomonas aeruginosa, wound

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Authors: A. T. Laiche, M. L. Tlil, Benine B., S. Bechoua

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The aim of this study is to isolate and select, from camel milk from El-Oued region in Algeria, a strains of lactic acid bacteria and possessing probiotic properties ; and to evaluate their potential effect on intestinal disorders in Wistar ratsmThe results relating to the selection of probiotic strains confirms that the Thermophilic streptococcus exhibits the best probiotic activity performance, with a resistance important to different degrees of pH and to bile salts, and a remarkable antibacterial activity and resistance to antibiotics compared to the other four isolated strains. In the in vivo study, diseases are induced in rats at the level of the digestive system, it was reported that the administration of Escherichia coli and castor oil caused an intestinal disorders. The microscopic observation of the histological section of the intestine showed a damaged intestinal structure and some symptoms of its irritation, including a decrease in the height of the villi and the presence of others destroyed cells, and after treatment with Streptococcus thermophilus, the microscopic observation of the cut of the histological section of the intestine showed almost complete disappearance of the mentioned symptoms, The dosage of the hematological parameters by complete blood count (CBC) is in agreement with the results of the histological sections.

Keywords: camel milk, probiotic, pathogenic bacteria, intestinal disorders, lactic acid bacteria

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Authors: Isabel Gouveia

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Antimicrobial textile materials may significantly reduce the risk of infections and because they are able to absorb substances from the skin and release therapeutic compounds to the skin, they can also find applications as complementary therapy of skin-diseases as part of standard management. Although functional textiles may be a promising area in skin disease/injury management, as part of standard management, few offer complementary treatment even though they are well known to reduce scratching and aiding emollient absorption, reducing infection, and alleviating pruritus. The reason for this may rely on the low quality of supporting evidence and negative effect that antimicrobial agents may exert on skin microbiome, as for example additional irritation of the vulnerable skin, and by causing resistant bacteria. Several antimicrobial agents have been tested in textiles: quaternary ammonium compounds, silver, polyhexamethylene-biguanides and triclosan have been used, with success. They have powerful bactericidal activity but the majority have a reduce spectrum of microbial inhibition and may cause skin irritation, ecotoxicity and bacteria resistance. Furthermore, the rising flow of strains resistant to last-resort antibiotics rekindles interest in alternative strategies. In this regard, new functional textiles incorporating highly specific antimicrobial agents towards pathogenic bacteria, are required. Recent research has been conducted on naturally occurring antimicrobials as novel alternatives to antibiotics. Conscious of this need our team firstly reported new approaches using L-cysteine and antimicrobial peptides (AMP). Briefly, we were able to develop different immobilization processes towards 6 Log Reduction against bacteria such as S. aureus and K. pneumoniae. Therefore, here we present several innovative antimicrobial textiles incorporating AMP and L-Cysteine which may open new avenues for the medical textiles market and biomaterials in general. Team references will be discussed as an overview and for comparison purposes in terms of potential therapeutic applications.

Keywords: Antimicrobials, Antimicrobial Textiles, Biomedical Textiles, Biomimetic surface functionalization

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Authors: Nura Brimo, Dilek Cokeliler Serdaroglu, Tansel Uyar, Busra Uysal, Elif Bahar Cakici, Miris Dikmen, Zerrin Canturk

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Background: A combination of antibiotics, including metronidazole (MET), ciprofloxacin (CIP), and minocycline (MINO), has been demonstrated to disinfect bacteria in necrotic teeth before regenerative processes. It has been presented clinically that antibiotic pastes may drive to possible stem cell death and difficulties in removing from the canal system, which can limit the regenerative procedure. This study was designed to (1) synthesize nanofibrous webs containing various concentrations of different medicaments (triple, double, and calcium hydroxide,Ca(OH)2), and (2) coat thiselectrospun fibrous gutta-percha (GP) cones. Methods: Poly(vinylpyrrolidone) (PVP)-based electrospun fibrous webs were processed with low medicaments concentrations. Scanning Electron Microscopy (SEM), Energy Dispersive X-Ray Spectroscopy (EDX), and X-Ray Photoelectron Spectroscopy (XPS) were carried out to investigate fiber morphology, antibiotic incorporation, and characterized GP-coated fibrous webs, respectively. The chemical and physical properties of dentine were carried out via Fourier Transform Infrared Spectroscopy (FTIR) and Nano-SEM, respectively. The antimicrobial properties of the different fibrous webs were assessed against various bacteria by direct nanofiber/bacteria contact. Cytocompatibility was measured by applying the MTT method. Results: The mean fiber diameter of the experiment groups of medicament-containing fibers ranged in the nm scale and was significantly smaller than PVP fibers. EDX analysis confirmed the presence of medicaments in the nanofibers. XPS analysis presented a complete coating of the fibers with GPs; FTIR and Nano-SEM showed no chemical and physical configuration of intracanal medicaments on the dentine surface. Meanwhile, nanofibrous webs led to a significant reduction in the percentage of viable bacteria compared with the negative control and PVP. Conclusion: Our findings suggest that TA-NFs, DA-NFs, and Cₐ(OH)₂)-NFs coated GP cones have significant potential in eliminating intracanal bacteria, cell-friendly behavior, and clinical usage features.

Keywords: drug delivery, drug carrier, electrospinning, nano/microfibers, regenerative endodontic, morphology

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Authors: Nada Verdel, Tomaz Rijavec, Albin Pintar, Ales Lapanje

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Objectives: White water circuits of woodfree paper mills contain suspended, dissolved, and colloidal particles, such as cellulose, starch, paper sizings, and dyes. By closing the white water circuits, these particles start to accumulate and affect the production. Due to high amount of organic matter that scavenge radicals and adsorbs onto catalyst surfaces, treatment of white water with photocatalysis is inappropriate. The most suitable approach should be bioaugmentation-assisted bioremediation. Accordingly, objectives were: - to isolate bacteria capable of degrading organic compounds used for the papermaking process - to select the most active bacteria for bioaugmentation. Status: The state-of-the-art of bioaugmentation of pulp and paper mill effluents is mostly based on biodegradation of lignin. Whereas in white water circuits of woodfree paper mills only papermaking compounds are present. As far as one can tell from the literature, the study on degradation activities of bacteria for all possible compounds of the papermaking process is a novelty. Methodology: The main parameters of the selected white water were systematically analyzed during a period of two months. Bacteria were isolated on selective media with particular carbon source. Organic substances used as carbon source either enter white water circuits as base paper or as recycled broke. The screening of bacterial activities for starch, cellulose, latex, polyvinyl alcohol, alkyl ketene dimers, and resin acids was followed by addition of lugol. Degraders of polycyclic aromatic dyes were selected by cometabolism tests; cometabolism is simultaneous biodegradation of two compounds, in which the degradation of the second compound depends on the presence of the first. The obtained strains were identified by 16S rRNA sequencing. Findings: 335 autochthonous strains were isolated on plates with selected carbon source. The isolated strains were selected according to degradation of the particular carbon source. The ultimate degraders of cationic starch, cellulose, and sizings are Pseudomonas sp. NV-CE12-CF and Aeromonas sp. NV-RES19-BTP. The most active strains capable of degrading azo dyes are Aeromonas sp. NV-RES19-BTP and Sphingomonas sp. NV-B14-CF. Klebsiella sp. NV-Y14A-BTP degrade polycyclic aromatic direct blue 15 and also yellow dye, Agromyces sp. NV-RED15A-BF and Cellulosimicrobium sp. NV-A4-BF are specialists for whitener and Aeromonas sp. NV-RES19-BTP is general degrader of all compounds. To the white water adapted bacteria were isolated and selected according to their degradation activities for particular organic substances. Mostly isolated bacteria are specialized to lower the competition in the microbial community. Degraders of readily-biodegradable compounds do not degrade recalcitrant polycyclic aromatic dyes and vice versa. General degraders are rare.

Keywords: bioaugmentation, biodegradation of azo dyes, cometabolism, smart wastewater treatment technologies

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Authors: Malory Jonata

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Chlordecone (CLD) is an organochlorine pesticide used between 1971 and 1993 in both Guadeloupe and Martinique for the control of banana black weevil. The bishomocubane structure which characterizes this chemical compound led to high stability in organic matter and high persistence in the environment. Recently, researchers found that CLD can be degraded by isolated bacteria consortiums and, particularly, by bacteria such as Citrobacter sp 86 and Delsulfovibrio sp 86. Actually, six transformation product families of CLD are known. Moreover, the latest discovery showed that CLD was disappearing faster than first predicted in highly contaminated soil in Guadeloupe. However, the toxicity of transformation products is still unknown, and knowledge has to be deepened on the degradation ways and chemical characteristics of chlordecone and its transformation products. Microbial fuel cells (MFC) are electrochemical systems that can convert organic matter into electricity thanks to electroactive bacteria. These bacteria can exchange electrons through their membranes to solid surfaces or molecules. MFC have proven their efficiency as bioremediation systems in water and soils. They are already used for the bioremediation of several organochlorine compounds such as perchlorate, trichlorophenol or hexachlorobenzene. In this study, a three-electrodes system, inspired by MFC, is used to try to degrade chlordecone using bacteria from a mangrove swamp in Martinique. As we know, some mangrove bacteria are electroactive. Furthermore, the CLD rate seems to decline in mangrove swamp sediments. This study aims to prove that electroactive bacteria from a mangrove swamp in Martinique can degrade CLD thanks to a three-electrodes bioelectrochemical system. To achieve this goal, the tree-electrodes assembly has been connected to a potentiostat. The substrate used is mangrove water and sediments sampled in the mangrove swamp of La Trinité, a coastal city in Martinique, where CLD contamination has already been studied. Electroactive biofilms are formed by imposing a potential relative to Saturated Calomel Electrode using chronoamperometry. Moreover, their comportment has been studied by using cyclic voltametry. Biofilms have been studied under different imposed potentials, several conditions of the substrate and with or without CLD. In order to quantify the evolution of CLD rates in the substrate’s system, gas chromatography coupled with mass spectrometry (GC-MS) was performed on pre-treated samples of water and sediments after short, medium and long-term contact with the electroactive biofilms. Results showed that between -0,8V and -0,2V, the three-electrodes system was able to reduce the chemical in the substrate solution. The first GC-MS analysis result of samples spiked with CLD seems to reveal decreased CLD concentration over time. In conclusion, the designed bioelectrochemical system can provide the necessary conditions for chlordecone degradation. However, it is necessary to improve three-electrodes control settings in order to increase degradation rates. The biological pathways are yet to enlighten by biologicals analysis of electroactive biofilms formed in this system. Moreover, the electrochemical study of mangrove substrate gives new informations on the potential use of this substrate for bioremediation. But further studies are needed to a better understanding of the electrochemical potential of this environment.

Keywords: bioelectrochemistry, bioremediation, chlordecone, mangrove swamp

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Authors: Tanju Teker, Yener Tekeli̇, Esra Karpuz

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Salvia species are known as medicinal plant and often used in public. The antimicrobial effects and essential oil composition of Salvia verticillata L. subsp. amasiaca were determined. The antimicrobial activity is determined by using disk diffusion method against two Gram-positive bacteria, two Gram-negative bacteria and one kind of yeast and essential oil composition was determined by GC - MS. As a result of antimicrobial analysis while sample has shown very strong antimicrobial activity against Staphylococcus aureus, moderately effective against Pseudomonas aeruginosa and low effective against Enterococcus faecalis, it has not shown antimicrobial activity against Escherichia coli and C. albicans. Trans-caryophyllene (% 35.07), germacrene-d (% 10.98) and caryopyllene oxide (% 5.81) are the main components of essential oil composition.

Keywords: salvia, medicinal plant, antimicrobial activity, essential oil

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Authors: Sidra Shaw, Sarenna Shaw, Chae Joon Lee, Irimpan Mathews, Eric Koehn

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One of the biggest public health concerns of our time is increasing antimicrobial resistance. As of 2019, the CDC has documented more than 2.8 million serious antibiotic resistant infections in the United States. Currently, antibiotic resistant infections are directly implicated in over 750,000 deaths per year globally. On our current trajectory, British economist Jim O’Neill predicts that by 2050, an additional 10 million people (about half the population of New York) will die annually due to drug resistant infections. As a result, new biochemical pathways must be targeted to generate next generation antibiotic drugs that will be effective against drug resistant bacteria. One enticing target is the biosynthesis of DNA within bacteria, as few drugs interrupt this essential life process. Thymidylate synthase enzymes are essential for life as they catalyze the synthesis of a DNA building block, 2′-deoxythymidine-5′-monophosphate (dTMP). In humans, the thymidylate synthase enzyme (TSase) has been shown to be distinct from the flavin-dependent thymidylate synthase (FDTS) produced by many pathogenic bacteria. TSase and FDTS have distinct structures and mechanisms of catalysis, which should allow selective inhibition of FDTS over human TSase. Currently, C. diff is one of the most antibiotic resistant bacteria, and no drugs that target thymine biosynthesis exist for C. diff. Here we present the initial biochemical characterization of FDTS from C. diff. Specifically, we examine enzyme kinetics and binding features of this enzyme to determine the nature of interaction with ligands/inhibitors and understand the molecular mechanism of catalysis. This research will provide more insight into the targetability of the C. diff FDTS enzyme for novel antibiotic drugs.

Keywords: flavin-dependent thymidylate synthase, FDTS, clostridioides difficile, C. diff, antibiotic resistance, DNA synthesis, enzyme kinetics, binding features

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Authors: Fatih Özogul, Nurten Toy, Yesim Özogul

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The influences of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on cadaverine and other biogenic amine production by Listeria monocytogenes and Staphylococcus aureus were investigated in lysine decarboxylase broth (LDB) using HPLC. Cell-free solutions were prepared from Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus acidophilus and Streptococcus thermophiles. Two different concentrations that were 50% and 25% CFS and the control without CFSs were prepared. Significant variations on biogenic amine production were observed in the presence of L. monocytogenes and S. aureus (P<0.05). The role of CFS on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine formation in control by L. monocytogenes and S. aureus were 500.9 and 948.1 mg/L, respectively while the CFSs of LAB induced 4-fold lower cadaverine production by L. monocytogenes and 7-fold lower cadaverine production by S. aureus. CFSs resulted in strong decreases in cadaverine and putrescine production by L. monocytogenes and S. aureus, although remarkable increases were observed for histamine, spermidine, spermine, serotonin, dopamine, tyramine, and agmatine, in the presence of LAB in lysine decarboxylase broth.

Keywords: cell-free solution, lactic acid bacteria, cadaverine, food borne-pathogen

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Authors: Garzon V., Bustos R., Salvador J. P., Marco M. P., Pinacho D. G.

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Bacterial resistance to antimicrobial treatment has increased significantly in recent years, making it a public health problem. Large numbers of bacteria are resistant to all or nearly all known antimicrobials, creating the need for the development of new types of antimicrobials or the use of “last line” antimicrobial drug therapies for the treatment of multi-resistant bacteria. Some of the chemical groups of antimicrobials most used for the treatment of infections caused by multiresistant bacteria in the clinic are Glycopeptide (Vancomycin), Polymyxin (Colistin), Lipopeptide (Daptomycin) and Carbapenem (Meropenem). Molecules that require therapeutic drug monitoring (TDM). Due to the above, a methodology based on nanobiotechnology based on an optical and electrochemical biosensor is being developed, which allows the evaluation of the plasmatic levels of some antimicrobials such as glycopeptide, polymyxin, lipopeptide and carbapenem quickly, at a low cost, with a high specificity and sensitivity and that can be implemented in the future in public and private health hospitals. For this, the project was divided into five steps i) Design of specific anti-drug antibodies, produced in rabbits for each of the types of antimicrobials, evaluating the results by means of an immunoassay analysis (ELISA); ii) quantification by means of an electrochemical biosensor that allows quantification with high sensitivity and selectivity of the reference antimicrobials; iii) Comparison of antimicrobial quantification with an optical type biosensor; iv) Validation of the methodologies used with biosensor by means of an immunoassay. Finding as a result that it is possible to quantify antibiotics by means of the optical and electrochemical biosensor at concentrations on average of 1,000ng/mL, the antibodies being sensitive and specific for each of the antibiotic molecules, results that were compared with immunoassays and HPLC chromatography. Thus, contributing to the safe use of these drugs commonly used in clinical practice and new antimicrobial drugs.

Keywords: antibiotics, electrochemical biosensor, optical biosensor, therapeutic drug monitoring

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Authors: Gina Zavaleta-Espejo, Segundo R. Jáuregui-Rosas, Fanny V. Samanamud-Moreno, José Saldaña Jiménez, Anibal Felix-Quintero, Víctor Montero-Del Aguila, Elsi Mejía-Uriarte

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Vicugnapacos "alpaca" fabrics are considered special for their finesse, and the garments in the textile market are very luxurious. It has many special characteristics such as antiallergic, soft, hygroscopic, among others. In this sense, the research aimed to evaluate the antimicrobial activity of alpaca fabrics functionalized with silver nanoparticles on the bacteria Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. For the functionalization of the fabrics, AgNO3 and different concentrations of trisodium citrate (TSC) 2, 6, and 10 mg. Tissue characterization was performed using Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The determination of the antimicrobial activity of the alpaca tissues was made by the Kirby-Bauer method with alpaca tissue discs functionalized with silver nanoparticles, an experimental design was made in completely randomized blocks with three treatments and a negative control with three repetitions. The results showed that inhibition halos were formed for both bacteria, therefore, the functionalized tissues have a high antimicrobial activity, whose mechanism of action is attributed to the free radicals (ROS) generated by the nanoparticles that cause oxidative damage to the bacteria. proteins and lipids of the bacterial cell wall.

Keywords: antimicrobial, animal fibers, fabrics, functionalization, trisodium citrate

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Authors: Moetazza M. Alshafei, Ahmed M. Mabrouk, Emtenan M. Hanafi, Manal M. Ramadan, Reda M. S. Korany, Seham S. Kassem, Dina Mostafa Mohammed

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Metabolic syndrome (MeS) is a major health problem with a high incidence of obese individuals worldwide. Increased related morbidity of diabetes, hypertension and fatty liver disease, and complicated cardiovascular disease are inevitable. Boswellia serrata gum (Bos) is a promising traditional medicinal plant; it has several pharmacological properties, including anti-inflammatory, antioxidant, and antilipase activities. Probiotics (Bac) supplements have good benefits on health and MeS, whether it is supplemented in combination with prebiotics or alone. Microencapsulation helps to mask unpalatable taste and odor and deliver active ingredients to targeted organs. Methodology MeS rat model was produced by feeding rats with a high fat, high CHO diet (HFD). Bos was extracted, and both Bos and the probiotic were microencapsulated with a spray drier. Female rats were divided into 5 groups (N8). HFD control, control normal receiving basic diet, HFD treated, from the start of the experiment, either with encapsulated Bos, Bac and Bos or Bac only, all treatments were received for eight weeks (after approval from NRC animal ethical committee). Serum was collected to analyze lipid profile, blood sugar, liver and kidney functions, antioxidants, leptin, and progesterone. Rat's organs and body fat were weighed and collected for histopathology. Statistical analysis was done by use of one way Anova test in the SPSS program. Results showed control of elevated body weight, lipid profile, and glucose levels as well as decrease of body fat index and improvement of histopathology of liver and heart, especially in combination. Conclusion: We concluded that both microencapsulated Bos and probiotics have a controlling effect on MeS parameters.

Keywords: metabolic syndrome, Boswellia serata, probiotic, micro-encapsulation, histopathology, liver steatosis

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Authors: Azizza Mala, Babo Fadlalla, Elnour Mohamed, Siran Wang, Junfeng Li, Tao Shao

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Sweet sorghum is considered one of the best plants for silage production and is now a more important feed crop in many countries worldwide. It is simple to ensile because of its high water-soluble carbohydrates (WSC) concentration and low buffer capacity. This study investigated the effect of adding Pediococcus acidilactici AZZ5 and Lactobacillus plantarum AZZ4 isolated from elephant grass on the fermentation quality of sweet sorghum silage. One commercial bacteria Lactobacillus Plantarum, Ecosyl MTD/1(C.B.)), and two strains were used as additives Pediococcus acidilactici (AZZ5), Lactobacillus plantarum subsp. Plantarum (AZZ4) at 6 log colony forming units (cfu)/g of fresh sweet sorghum grass in laboratory silos (1000g). After 15, 30, and 60 days, the silos for each treatment were opened. All of the isolated strains enhanced the silage quality of sweet sorghum silage compared to the control, as evidenced by significantly (P < 0.05) lower ammonia nitrogen (NH3-N) content and undesirable microbial counts, as well as greater lactic acid (L.A.) contents and lactic acid/acetic acid (LA/AA) ratios. In addition, AZZ4 performed better than all other inoculants during ensiling, as evidenced by a significant (P < 0.05) reduction in pH and ammonia-N contents and a significant increase in lactic acid contents.

Keywords: fermentation, lactobacillus plantarum, lactic acid bacteria, pediococcus acidilactic, sweet sorghum

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Authors: Lígia Pimentel, Miguel Pereira, Manuela Pintado

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Wheat germ is a by-product of wheat flour refining. Despite this by-product being a source of proteins, lipids, fibres and complex carbohydrates, and consequently a valuable ingredient to be used in Food Industry, only few applications have been studied. The main goal of this study was to assess the potential prebiotic effect of natural wheat germ. The prebiotic potential was evaluated by in vitro assays with individual microbial strains (Lactobacillus paracasei L26 and Lactobacillus casei L431). A simulated model of the gastrointestinal digestion was also used including the conditions present in the mouth (artificial saliva), oesophagus–stomach (artificial gastric juice), duodenum (artificial intestinal juice) and ileum. The effect of natural wheat germ and wheat germ after digestion on the growth of lactic acid bacteria was studied by growing those microorganisms in de Man, Rogosa and Sharpe (MRS) broth (with 2% wheat germ and 1% wheat germ after digestion) and incubating at 37 ºC for 48 h with stirring. A negative control consisting of MRS broth without glucose was used and the substrate was also compared to a commercial prebiotic fructooligosaccharides (FOS). Samples were taken at 0, 3, 6, 9, 12, 24 and 48 h for bacterial cell counts (CFU/mL) and pH measurement. Results obtained showed that wheat germ has a stimulatory effect on the bacteria tested, presenting similar (or even higher) results to FOS, when comparing to the culture medium without glucose. This was demonstrated by the viable cell counts and also by the decrease on the medium pH. Both L. paracasei L26 and L. casei L431 could use these compounds as a substitute for glucose with an enhancement of growth. In conclusion, we have shown that wheat germ stimulate the growth of probiotic lactic acid bacteria. In order to understand if the composition of gut bacteria is altered and if wheat germ could be used as potential prebiotic, further studies including faecal fermentations should be carried out. Nevertheless, wheat germ seems to have potential to be a valuable compound to be used in Food Industry, mainly in the Bakery Industry.

Keywords: by-products, functional ingredients, prebiotic potential, wheat germ

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Authors: Navid Mosallaei, Mahmoud Reza Jaafari, Mohammad Yahya Hanafi-Bojd, Shiva Golmohammadzadeh, Bizhan Malaekeh-Nikouei

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Background: Docetaxel (DTX), a potent anticancer drug derived from the European yew tree, is effective against various human cancers by inhibiting microtubule depolymerization. Solid lipid nanoparticles (SLNs) have gained attention as drug carriers for enhancing drug effectiveness and safety. SLNs, submicron-sized lipid-based particles, can passively target tumors through the "enhanced permeability and retention" (EPR) effect, providing stability, drug protection, and controlled release while being biocompatible. Methods: The SLN formulation included biodegradable lipids (Compritol and Precirol), hydrogenated soy phosphatidylcholine (H-SPC) as a lipophilic co-surfactant, and Poloxamer 188 as a non-ionic polymeric stabilizer. Two SLN preparation techniques, probe sonication and microemulsion, were assessed. Characterization encompassed SLNs' morphology, particle size, zeta potential, matrix, and encapsulation efficacy. In-vitro cytotoxicity and cellular uptake studies were conducted using mouse colorectal (C-26) and human malignant melanoma (A-375) cell lines, comparing SLN-DTX with Taxotere®. In-vivo studies evaluated tumor inhibitory efficacy and survival in mice with colorectal (C-26) tumors, comparing SLNDTX withTaxotere®. Results: SLN-DTX demonstrated stability, with an average size of 180 nm and a low polydispersity index (PDI) of 0.2 and encapsulation efficacy of 98.0 ± 0.1%. Differential scanning calorimetry (DSC) suggested amorphous encapsulation of DTX within SLNs. In vitro studies revealed that SLN-DTX exhibited nearly equivalent cytotoxicity to Taxotere®, depending on concentration and exposure time. Cellular uptake studies demonstrated superior intracellular DTX accumulation with SLN-DTX. In a C-26 mouse model, SLN-DTX at 10 mg/kg outperformed Taxotere® at 10 and 20 mg/kg, with no significant differences in body weight changes and a remarkably high survival rate of 60%. Conclusion: This study concludes that SLN-DTX, prepared using the probe sonication, offers stability and enhanced therapeutic effects. It displayed almost same in vitro cytotoxicity to Taxotere® but showed superior cellular uptake. In a mouse model, SLN-DTX effectively inhibited tumor growth, with 10 mg/kg outperforming even 20 mg/kg of Taxotere®, without adverse body weight changes and with higher survival rates. This suggests that SLN-DTX has the potential to reduce adverse effects while maintaining or enhancing docetaxel's therapeutic profile, making it a promising drug delivery strategy suitable for industrialization.

Keywords: docetaxel, Taxotere®, solid lipid nanoparticles, enhanced permeability and retention effect, drug delivery, cancer chemotherapy, cytotoxicity, cellular uptake, tumor inhibition

Procedia PDF Downloads 49

Authors: Maria Manzoor, Iram Gul, Muhammad Arshad

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Bacterial strains were isolated from contaminated soil collected from Rawalpindi and Islamabad. The strains were investigated for lead resistance and their effect on Pb solubility and PGPR activity. Incubation experiments were carried for inoculated and unoculated soil containing different levels of Pb. Results revealed that few stains (BTM-4, BTM-11, BTM-14) were able to tolerate Pb up to 600 mg L-1, whereas five strains (BTM-3, BTM-6, BTM-10, BTM-21 and BTM-24) showed significant increase in solubility of Pb when compared to all other strains and control. The CaCl2 extractable Pb was increased by 13.6, 6.8, 4.4 and 2.4 folds compared to un-inoculated control soil at increased soil Pb concentration (500, 1000, 1500 and 200 mg kg-1, respectively). The selected bacterial strains (11) were further investigated for plant growth promotion activity through PGPR assays including. Germination and root elongation assays were also conducted under elevated metal concentration in controlled conditions to elucidate the effects of microbial strains upon plant growth and development. The results showed that all the strains tested in this study, produced significantly varying concentrations of IAA, siderophores and gibberellic acid along with ability to phosphorus solubilization index (PSI). The results of germination and root elongation assay further confirmed the beneficial role of the microbial strains in elevating metal stress through PGPR activity. Among all tested strains, BTM-10 significantly improved plant growth. 1.3 and 2.7 folds increase in root and shoot length was observed when compared to control. Which may be attributed to presence of important plant growth promoting enzymes (IAA 74.6 μg/ml; GA 19.23 μg/ml; Sidrophore units 49% and PSI 1.3 cm). The outcome of this study indicates that these Pb tolerant and solubilizing strains may have the potential for plant growth promotion under metal stress and can be used as mediator when coupled with heavy metal hyperaccumulator plants for phytoremediation of Pb contaminated soil.

Keywords: Pb resistant bacteria, Pb mobilizing bacteria, Phytoextraction of Pb, PGPR activity of bacteria

Procedia PDF Downloads 191

Authors: Sonja Djilas, Aleksandra Velićanski, Dragoljub Cvetković, Siniša Markov, Eva Lončar, Vesna Tumbas Šaponjac, Milica Vinčić

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Due to high content of bioactive compounds, sour cherry possesses antioxidant and antimicrobial activity. Additionally, waste material from industrial processing of sour cherry is also a good source of bioactive compounds. The aim of this study was to screen the antimicrobial activity and determine the minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC) of sour cherry pomace extract. Tested strains were Gram-negative bacteria (Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14028 and wild isolates Escherichia coli and Salmonella sp.), Gram-positive bacteria (Staphylococcus aureus ATCC 11632, Bacillus cereus ATCC 10876 and wild isolates Staphylococcus saprophyticus and Bacillus sp.) and yeasts (Saccharomyces cerevisiae 112, Hefebank Weihenstephan and Candida albicans ATCC 10231). Antimicrobial activity was tested by disc-diffusion method and agar-well diffusion method. MIC and MBC were determined by microdilution method. Screening tests showed that Gram-negative bacteria were resistant to tested extract, with exception of Salmonella typhimurium and Salmonella sp. for which only zones of reduced growth appeared. However, Gram-positive bacteria were more sensitive where the highest clear zones appeared with 100 µl of extract applied. There was no activity against tested yeasts. MIC and MBC values were in the range 3.125-37.5 mg/ml and 6.25-100 mg/ml, respectively. The most susceptible strain was Staphylococcus aureus while the most resistant was Bacillus sp. where MBC was not found in tested concentration range. Sour cherry pomace possesses high antibacterial potential, which indicates that this waste material is a promising source of bioactive compounds and could be used as a functional food ingredient.

Keywords: antimicrobial activity, sour cherry, pomace, bioactive compounds

Procedia PDF Downloads 305

Authors: Hakimullah Hakim, Chiharu Toyofuku, Mari Ota, Mayuko Suzuki, Miyuki Komura, Masashi Yamada, Md. Shahin Alam, Natthanan Sangsriratanakul, Dany Shoham, Kazuaki Takehara

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Disinfectants and their application are essential part of infection control strategies and enhancement of biosecurity at farms, worldwide. Alkaline agents are well known for their strong and long term antimicrobial capacities and most frequently are applied at farms for control and prevention of biological hazards. However, inadequate information regarding such materials’ capacities to inactivate pathogens and their improper applications fail farmers to achieve the mentioned goal. Thus, this requires attention to further evaluate their efficacies, under different conditions and in different ways. Here in this study we evaluated bactericidal efficacies of food additive grade of calcium hydroxide (FdCa(OH)2) powder derived from natural calcium carbonates obtained from limestone (Fine Co., Ltd., Tokyo, Japan), and bioceramic powder (BCX) derived from chicken feces at pH 13 (NMG environmental development Co., Ltd., Tokyo, Japan), for their efficacies to inactivate bacteria in feces. [Materials & Methods] Chicken feces were inoculated by 100 µl Escherichia coli and Salmonella Infantis in falcon tubes, individually, then FdCa(OH)2 or BCX powders were individually added to make final concentration of 0, 5, 10, 20 and 30% (w/w) in total weight of 0.5g, followed by properly mixing and incubating at room temperature for certain periods of time, in a dark place. Afterwards, 10 ml 1M Tris-HCl (pH 7.2) was added onto them to reduce their pH, in order to stop powders’ activities and to harvest the remained viable bacteria, whereas using normal medium or dW2 to recover bacteria increases the mixture pH, and as a result bacteria would be inactivated soon; therefore, the latter practice brings about incorrect and misleading results. Samples were then inoculated on DHL agar plates in order to calculate colony forming units (CFU)/ml of viable bacteria. [Results and Discussion] FdCa(OH)2 powder at 10% and 5% required 3 hr and 6 hr exposure times, respectively, while BCX powder at 20% concentrations required 6 hr exposure time to kill the mentioned bacteria in feces down to lower than detectable level (≤ 3.6 log10 CFU/ml). This study confirmed capacities of FdCa(OH)2 and BCX powders to inactivate bacteria in feces, and both materials are environment friendly materials, with no risk to human or animal’s health. This finding helps farmers to properly apply alkaline agents in appropriate concentrations and exposure times in their farms, in order to prevent and control infectious diseases outbreaks and to enhance biosecurity. Finally, this finding may help farmers to implement better strategies for infections control in their livestock farms.

Keywords: bacterial inactivation, bioceramic, biosecurity at livestock farms, chicken feces

Procedia PDF Downloads 411

Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

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Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

Procedia PDF Downloads 147

Authors: Mohammadhosein Rahimi, Mohammad Raouf Hosseini, Mehran Bakhshi, Alireza Baghbanan

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Silver ions (Ag⁺) and their compounds are consequentially toxic to microorganisms, showing biocidal effects on many species of bacteria. Silver-phosphate (or silver orthophosphate) is one of these compounds, which is famous for its antimicrobial effect and catalysis application. In the present study, a green method was presented to synthesis silver-phosphate nanoparticles using Sporosarcina pasteurii. The composition of the biosynthesized nanoparticles was identified as Ag₃PO₄ using X-ray Diffraction (XRD) and Energy Dispersive Spectroscopy (EDS). Also, Fourier Transform Infrared (FTIR) spectroscopy showed that Ag₃PO₄ nanoparticles was synthesized in the presence of biosurfactants, enzymes, and proteins. In addition, UV-Vis adsorption of the produced colloidal suspension approved the results of XRD and FTIR analyses. Finally, Transmission Electron Microscope (TEM) images indicated that the size of the nanoparticles was about 20 nm.

Keywords: bacteria, biosynthesis, silver-phosphate, Sporosarcina pasteurii, nanoparticle

Procedia PDF Downloads 421

Authors: Uftah Ali M. Shushni, Viola Stuppec, Ulrike Lindequist

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Because of the evolving resistance of microorganisms to existing antibiotics, there is an increasing need for new antibiotics not only in human but also in veterinary medicine. As part of our ongoing work on the secondary metabolites produced by marine fungi, the organic extract of the culture filtrate of an Ascomycetous fungus, which was found on driftwood collected from the coast of the Greifswalder Bodden, Baltic Sea, Germany displayed antimicrobial activity against some fish and human pathogenic bacteria. Bioactivity-guided column chromatographic separation led to the isolation of 6-Deoxybostrycoidin. The structure was determined from the interpretation of spectroscopic data (UV, MS, and NMR). 6-Deoxybostrycoidin exhibited in vitro activity against Bacillus subtilis, Staphylococcus aureus and Flexibacter maritimus with minimal inhibitory concentrations of 25, 12.5 and 12.5 μg/ml respectively.

Keywords: marine fungi, fish pathogenic bacteria, microorganism, medicine

Procedia PDF Downloads 501

Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

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Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

Procedia PDF Downloads 372

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

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Urinary tract infections (UTIs) are considered to be the most common bacterial infections worldwide, which are caused mainly by Escherichia (E.) coli (about 80%). Klebsiella pneumoniae (about 10%) and Pseudomonas aeruginosa (about 6%). Although antibiotics are considered as the most effective treatment for bacterial infectious diseases, unfortunately, most of the bacteria already have developed resistance to the majority of the commonly available antibiotics. Therefore, it is crucial to identify the infecting bacteria and to determine its susceptibility to antibiotics for prescribing effective treatment. Classical methods are time consuming, require ~48 hours for determining bacterial susceptibility. Thus, it is highly urgent to develop a new method that can significantly reduce the time required for determining both infecting bacterium at the species level and diagnose its susceptibility to antibiotics. Fourier-Transform Infrared (FTIR) spectroscopy is well known as a sensitive and rapid method, which can detect minor molecular changes in bacterial genome associated with the development of resistance to antibiotics. The main goal of this study is to examine the potential of FTIR spectroscopy, in tandem with machine learning algorithms, to identify the infected bacteria at the species level and to determine E. coli susceptibility to different antibiotics directly from patients' urine in about 30minutes. For this goal, 1600 different E. coli isolates were isolated for different patients' urine sample, measured by FTIR, and analyzed using different machine learning algorithm like Random Forest, XGBoost, and CNN. We achieved 98% success in isolate level identification and 89% accuracy in susceptibility determination.

Keywords: urinary tract infections (UTIs), E. coli, Klebsiella pneumonia, Pseudomonas aeruginosa, bacterial, susceptibility to antibiotics, infrared microscopy, machine learning

Procedia PDF Downloads 136

Authors: H. K. Ratre, M. Roy, S. Roy, M. S. Parmar, V. Bhagat

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Mastitis is a common problem of dairy industries. Reduction in milk production and an irreparable damage to the udder associated with the disease are common causes of culling of dairy cows. Milk from infected animals is not suitable for drinking and for making different milk products. So, it has a major economic importance in dairy cattle. The aims of this study were to investigate the bacteriological panorama in milk from udder quarters with subclinical mastitis and to carried out for the molecular characterization of the major isolated organisms, from subclinical mastitis-affected cows in and around Durg and Rajnandgaon district of Chhattisgarh. Isolation and identification of bacteria from the milk samples of subclinical mastitis-affected cows were done by standard and routine culture procedures. A total of 78 isolates were obtained from cows and among the various bacteria isolated, Staphylococcus spp. occupied prime position with occurrence rate of 51.282%. However, other bacteria isolated includeStreptococcus spp. (20.512%), Micrococcus spp. (14.102%), E. coli (8.974%), Klebsiela spp. (2.564%), Salmonella spp. (1.282%) and Proteus spp. (1.282%). Staphylococcus spp. was isolated as the major causative agent of subclinical mastitis in the studied area. Molecular characterization of Staphylococus aureusisolates was done for genetic expression of the virulence genes like ‘nuc’ encoding thermonucleaseexoenzyme, coa and spa by PCR amplification of the respective genes in 25 Staphylococcus isolates. In the present study, 15 isolates (77.27%) out of 20 coagulase positive isolates were found to be genotypically positive for ‘nuc’ where as 20 isolates (52.63%) out of 38 CNS expressed the presence of the same virulence gene. In the present study, three Staphylococcus isolates were found to be genotypically positive for coa gene. The Amplification of the coa gene yielded two different products of 627, 710 bp. The amplification of the gene segment encoding the IgG binding region of protein A (spa) revealed a size of 220 and 253bp in twostaphylococcus isolates. The X-region binding of the spa gene produced an amplicon of 315 bp in one Staphylococcal isolates. Staphylococcus aureus was found to be major isolate (51.28%) responsible for causing subclinical mastitis in cows which also showed expression of virulence genesnuc, coa and spa.

Keywords: mastitis, bacteria, characterization, expression, gene

Procedia PDF Downloads 189

Authors: Mihriban Korukluoglu, Goksen Arik, Cagla Erdogan, Selen Kocakoglu

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Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Keywords: antagonistic effect, kefir, lactic acid bacteria (LAB), synergistic, yeast

Procedia PDF Downloads 251

Authors: Hardik Goswami, Gayatri Swarnakar

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-Mastitis disease has been known as one of the most costly diseases of dairy cattle and observed as an inflammatory disease of cow and buffalo udder. Mastitis badly affected animal health, quality of milk and economics of milk production along with cause’s great economic loss. Bacteria have been representing the most common etiological agents of mastitis. The antibiotic sensitivity test was important to attain accurate treatment of mastitis. The aim of present research work was to explore prevalence and antibiotic susceptibility pattern of bacterial isolates recovered from cow and buffalo clinical mastitis milk sample. During the period of April 2010 to April 2014, total 1487 clinical mastitis milk samples of cow and buffalo were tested to check the prevalence of mastitis causing bacterial isolates. Milk samples were collected aseptically from the udder at the time of morning milking. The most prevalent bacterial isolates were Staphylococcus aureus (24.34%) followed by coliform bacteria (15.87%), coagulase negative Staphylococcus aureus (13.85%), non-coliform bacteria (13.05%), mixed infection (12.51%), Streptococcus spp. (10.96%). Out of 1487, 140 (9.42%) mastitis milk samples showed no growth on culture media. Identification of bacteria made on the basis of Standard Microbial features and procedures. Antibiotic susceptibility of bacterial isolates was investigated by Kirby-Bauer disk diffusion method. In vitro Antibiotic susceptibility test of bacterial isolates revealed higher sensitivity to Gentamicin (74.6%), Ciprofloxacin (62.1%) and Amikacin (59.4%). The lower susceptibility was shown to Amoxicillin (21.6%), Erythromycin (26.4%) and Ceftizoxime (29.9%). Antibiotic sensitivity pattern revealed Gentamicin are the possible effective antibiotic against the major prevalent mastitis pathogens. Present research work would be helpful in increase production, quality and quantity of milk, increase annual income of dairy owners and improve health of cow and buffaloes.

Keywords: antibiotic, buffalo, cow, mastitis, prevalence

Procedia PDF Downloads 373

Authors: Morteza Haghi, Cigdem Yilmazbas, Ayse Zeynep Uysal, Melisa Tepedelen, Gozde Turkoz Bakirci

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Today, the increase in plastic waste accumulation is an inescapable consequence of environmental pollution; the disposal of these wastes has caused a significant problem. Variable methods have been utilized; however, biodegradation is the most environmentally friendly and low-cost method. Accordingly, the present study aimed to isolate the bacteria capable of biodegradation of plastics. In doing so, we applied the liquid carbon-free basal medium (LCFBM) prepared with deionized water for the isolation of bacterial species obtained from soil samples taken from the Izmir Menemen region. Isolates forming biofilms on plastic were selected and named (PLB3, PLF1, PLB1B) and subjected to a degradation test. FTIR analysis, 16s rDNA amplification, sequencing, identification of isolates were performed. Finally, at the end of the process, a mass loss of 16.6% in PLB3 isolate and 25% in PLF1 isolate was observed, while no mass loss was detected in PLB1B isolate. Only PLF1 and PLB1B created transparent zones on plastic texture. Considering the FTIR result, PLB3 changed plastic structure by 13.6% and PLF1 by 17%, while PLB1B did not change the plastic texture. According to the 16s rDNA sequence analysis, FLP1, PLB1B, and PLB3 isolates were identified as Streptomyces albogriseolus, Enterobacter cloacae, and Klebsiella pneumoniae, respectively.

Keywords: polyethylene, biodegradation, bacteria, 16s rDNA, FTIR

Procedia PDF Downloads 169