Search results for: pathogenic microorganisms

Authors: A. Ribeiro, N. Valério, C. Vilarinho, J. Araujo, J. Carvalho

Abstract:

Res2ValHUM project involves institutions from the Spanish Autonomous Region of Galicia and the north of Portugal (districts of Porto and Braga) and has as overall objectives of promotion of composting as an process for the correct managing of organic waste, valorization of compost in different fields or applications for the constitution of products with high added value, reducing of raw materials losses, and reduction of the amount of waste throw in landfills. Three main actions were designed to achieve the objectives: development of a management tool to improve collection and residue channeling for composting, sensibilization of the population for composting and characterization of the chemical and biological properties of compost and humic and fulvic substances to envisage high-value applications of compost. Here we present the cooperative activity of Galician and northern Portuguese institutions to valorize organic waste in both regions with common socio-economic characteristics and residue management problems. Results from the creation of the resource manage tool proved the existence of a large number of agricultural wastes that could be valorized. In the North of Portugal, the wastes from maize, oats, potato, apple, grape pomace, rye, and olive pomace can be highlighted. In the Autonomous Region of Galicia the wastes from maize, wheat, potato, apple, and chestnuts can be emphasized. Regarding the isolation and identification of microbial consortia from compost samples, results proved microorganisms belong mainly to the genus Bacillus spp. Among all the species identified in compost samples, Bacillus licheniformis can be highlighted in the production of humic and fulvic acids.

Keywords: agricultural wastes, Bacillus licheniformis, Bacillus spp., humic-acids, fulvic-acids

Procedia PDF Downloads 99

Authors: Vikash Yadav, Ashish Arora

Abstract:

Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.

Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator

Procedia PDF Downloads 52

Authors: Mohammad Hossein Karimi Darvanjooghi, Sara Magdouli, Satinder Kaur Brar

Abstract:

It has been reported that the microorganism’s attachment to the surfaces of ore samples is a key factor that influences the biooxidation in pretreatment for recovery of gold in sulfide-bearing ores. In this research, the implementation of D-(+)-Sucrose on the biooxidation of ore samples were studied in a semi-pilot experiment. The experiments were carried out in five separate jacketed columns (1 m height and 6 cm diameter) at a constant temperature of 37.5 ̊C and saturated humidity. The airflow rate and recycling solution flow rate were studied in the research and the optimum operating condition were reported. The ore sample (0.49 ppm gold grade) was obtained from the Hammond Reef mine site containing 15 wt.% of pyrite which included 98% of gold according to the results of micrograph images. The experiments were continued up to 100 days while air flow rates were chosen to be 0.5, 1, 1.5, 2, and 3 lit/min and the recycling solution (Containing 9K media and 0.4 wt.% D-(+)-Sucrose) flow rates were kept 5, 8, 15 ml/hr. The results indicated that the addition of D-(+)-Sucrose increased the bacterial activity due to the overproduction of extracellular polymeric substance (EPS) up to 95% and for the condition that the recycling solution and air flow rate were chosen to be 8 ml/hr and 2 lit/min, respectively, the maximum pyrite dissolution of 76% was obtained after 60 days. The results indicated that for the air flow rates of 0.5, 1, 1.5, 2, and 3 lit/min the ratio of daily pyrite dissolution per daily solution lost were found to be 0.025, 0.033, 0.031, 0.043, and 0.009 %-pyrite dissolution/ml-lost. The implementation of this microorganisms and the addition of D-(+)-Sucrose will enhance the efficiency of gold recovery through faster biooxidation process and leads to decrease in the time and energy of operation toward desired target; however, still other parameters including particle size distribution, agglomeration, aeration design, chemistry of recycling solution need to be controlled and monitored for reaching the optimum condition.

Keywords: column tests, biooxidation, gold recovery, Ferroplasma acidophilum, optimization

Procedia PDF Downloads 47

Authors: Mesih Kocamuftuoglu, Gonca Ozan, Enver Ozan, Nalan Kaya, Sema Temizer Ozan

Abstract:

Metabolic Syndrome, one of the common metabolic disorder, occurs with co-development of insulin resistance, obesity, dislipidemia and hypertension problems. Insulin resistance appears to play a pathogenic role in the metabolic syndrome. Also, there is a relationship between insulin resistance and infertility as known. Turmeric (Curcuma longa L.) a polyphenolic chemical is widely used for its coloring, flavoring, and medicinal properties, and exhibits a strong antioxidant activity. In this study, we assess the effects of turmeric on rat uterine tissue in metabolic syndrome model induced by high fructose diet. Thirty-two adult female Wistar rats weighing 220±20 g were randomly divided into four groups (n=8) as follows; control, fructose, turmeric, and fructose plus turmeric. Metabolic syndrome was induced by fructose solution 20% (w/v) in tap water, and turmeric (C.Longa) administered at the dose of 80 mg/kg body weight every other day by oral gavage. After the experimental period of 8 weeks, rats were decapitated, serum and uterine tissues were removed. Serum lipid profile, glucose, insülin levels were measured. Uterine tissues were fixed for histological analyzes. The uterine tissue sections were stained with hematoxylin-eosin (H & E) stain, then examined and photographed on a light microscope (Novel N-800Mx20). As a result, fructose consumption effected serum lipids, insulin levels, and insulin resistance significantly. Endometrium and myometrium layers were observed in normal structure in control group of uterine tissues. Perivascular edema, peri glandular fibrosis, and inflammatory cell increase were detected in fructose group. Sections of the fructose plus turmeric group showed a significant improvement in findings when compared to the fructose group. Turmeric group cell structures were observed similar with the control group. These results demonstrated that high-fructose consumption could change the structure of the uterine tissue. On the other hand, turmeric administration has beneficial effects on uterine tissue at that dose and duration when administered with fructose.

Keywords: metabolic syndrome, rat, turmeric, uterus

Procedia PDF Downloads 156

Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc

Abstract:

Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.

Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation

Procedia PDF Downloads 298

Authors: Ali Ghorbanipour

Abstract:

The aim of this study is to investigate the drug resistance pattern and the value of the MIC (minimum inhibitory concentration)method to reduce the impact of infectious diseases and the slow development of resistance. Method: The study was conducted on clinical specimens collected between 2018 to 2021. identification of isolates and antibiotic susceptibility testing were performed using conventional biochemical tests. Antibiotic resistance was determined using kibry-Bauer disk diffusion and MIC by E-test methods comparative with microdilution plate elisa method. Results were interpreted according to CLSI. Results: Out of 249600 different clinical specimens, 18720 different pathogenic bacteria by overall detection ratio 7.7% were detected. Among pathogen bacterial were Gram negative bacteria (70%,n=13000) and Gram positive bacteria(30%,n=5720).Medically relevant gram-negative bacteria include a multitude of species such as E.coli , Klebsiella .spp , Pseudomonas .aeroginosa , Acinetobacter .spp , Enterobacterspp ,and gram positive bacteria Staphylococcus.spp , Enterococcus .spp , Streptococcus .spp was isolated . Conclusion: Our results highlighted that the resistance ratio among Gram Negative bacteria and Gram positive bacteria with different infection is high it suggest constant screening and follow-up programs for the detection of antibiotic resistance and the value of MIC drug susceptibility reporting that provide a new way to the usage of resistant antibiotic in combination with other antibiotics or accurate weight of antibiotics that inhibit or kill bacteria. Evaluation of wrong medication in the expansion of resistance and side effects of over usage antibiotics are goals. Ali ghorbanipour presently working as a supervision at the microbiology department of Massoud medical laboratory. Iran. Earlier, he worked as head department of pulmonary infection in firoozgarhospital, Iran. He received master degree in 2012 from Fergusson College. His research prime objective is a biologic wound dressing .to his credit, he has Published10 articles in various international congresses by presenting posters.

Keywords: antimicrobial profile, MIC & MBC Method, microplate antimicrobial assay, E-test

Procedia PDF Downloads 110

Authors: Ozan Kahraman, Hao Feng

Abstract:

Several models such as Weibull, Modified Gompertz, Biphasic linear, and Log-logistic models have been proposed in order to describe non-linear inactivation kinetics and used to fit non-linear inactivation data of several microorganisms for inactivation by heat, high pressure processing or pulsed electric field. First-order kinetic parameters (D-values and z-values) have often been used in order to identify microbial inactivation by non-thermal processing methods such as ultrasound. Most ultrasonic inactivation studies employed first-order kinetic parameters (D-values and z-values) in order to describe the reduction on microbial survival count. This study was conducted to analyze the E. coli O157:H7 inactivation data by using five microbial survival models (First-order, Weibull, Modified Gompertz, Biphasic linear and Log-logistic). First-order, Weibull, Modified Gompertz, Biphasic linear and Log-logistic kinetic models were used for fitting inactivation curves of Escherichia coli O157:H7. The residual sum of squares and the total sum of squares criteria were used to evaluate the models. The statistical indices of the kinetic models were used to fit inactivation data for E. coli O157:H7 by MTS at three temperatures (40, 50, and 60 0C) and three pressures (100, 200, and 300 kPa). Based on the statistical indices and visual observations, the Weibull and Biphasic models were best fitting of the data for MTS treatment as shown by high R2 values. The non-linear kinetic models, including the Modified Gompertz, First-order, and Log-logistic models did not provide any better fit to data from MTS compared the Weibull and Biphasic models. It was observed that the data found in this study did not follow the first-order kinetics. It is possibly because of the cells which are sensitive to ultrasound treatment were inactivated first, resulting in a fast inactivation period, while those resistant to ultrasound were killed slowly. The Weibull and biphasic models were found as more flexible in order to determine the survival curves of E. coli O157:H7 treated by MTS on apple-carrot juice.

Keywords: Weibull, Biphasic, MTS, kinetic models, E.coli O157:H7

Procedia PDF Downloads 343

Authors: Lígia Pimentel, Miguel Pereira, Manuela Pintado

Abstract:

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

Procedia PDF Downloads 461

Authors: Tooba N. Shamsi, Romana Perveen, Sadaf Fatima

Abstract:

Protease Inhibitors (PIs) are widespread in nature, produced by animals, plants and microorganisms. They play vital role in various biological activities by keeping a check on activity of proteases. Present study aims to investigate antioxidant and anti-inflammatory properties of PPI from Cajanus cajan (CCTI) and Phaseolus limensis (LBTI). PPI was purified from C. cajan (PUSA-992) by ammonium sulfate precipitation followed by ion exchange chromatography. The anti-oxidant activity was analyzed by two most common radical scavenging assays of FRAP (ferric reducing antioxidant power) and DPPH (1,1- diphenyl-2-picrylhydrazyl). Also, in-vitro anti-inflammatory activity was evaluated using albumin denaturation assay and membrane stabilization assay at different concentrations. Ascorbic acid and aspirin were used as a standards for antioxidant and anti-inflammatory assays respectively. The PPIs were also checked for antimicrobial activity against a number of bacterial strains. The CCTI and LBTI showed DPPH radical scavenging activity in a concentration–dependent manner with IC50 values 544 µg/ml and 506 µg/ml respectively comparative to ascorbic acid which was 258 µg/ml. Following FRAP assay, it was evaluated that LBTI had 87.5% and CCTI showed 84.4% antioxidant activity, taking value of standard ascorbic acid to be 100%. The PPIs also showed in-vitro anti‐inflammatory activity by inhibiting the heat induced albumin denaturation with IC50 values of 686 µg/ml and 615 µg/ml for CCTI and LBTI respectively compared to the standard (aspirin) which was 70.8 µg/ml. Red blood cells membrane stabilization with IC50 values of 641 µg/ml and 587 µg/ml for CCTI and LBTI respectively against aspirin which showed IC50 value of 70.4 µg/ml. PPIs showed antibacterial activity against 7 known strains while there was apparently no action against fungi.

Keywords: Cajanus cajan, Phaseolus limensis, Lima beans, protein protease inhibitor, antioxidant, anti-inflammatory, antimicrobial activity

Procedia PDF Downloads 269

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

Abstract:

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

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

Procedia PDF Downloads 245

Authors: Vaishali Patil, Neeraj Masand

Abstract:

In older people, Alzheimer’s disease (AD) is turning out to be a lethal disease. According to the amyloid hypothesis, aggregation of the amyloid β–protein (Aβ), particularly its 42-residue variant (Aβ42), plays direct role in the pathogenesis of AD. Aβ is generated through sequential cleavage of amyloid precursor protein (APP) by β–secretase (BACE) and γ–secretase (GS). Thus in the treatment of AD, γ-secretase modulators (GSMs) are potential disease-modifying as they selectively lower pathogenic Aβ42 levels by shifting the enzyme cleavage sites without inhibiting γ–secretase activity. This possibly avoids known adverse effects observed with complete inhibition of the enzyme complex. Virtual screening, via drug-like ADMET filter, QSAR and molecular docking analyses, has been utilized to identify novel γ–secretase modulators with sulphonamide nucleus. Based on QSAR analyses and docking score, some novel analogs have been synthesized. The results obtained by in silico studies have been validated by performing in vivo analysis. In the first step, behavioral assessment has been carried out using Scopolamine induced amnesia methodology. Later the same series has been evaluated for neuroprotective potential against the oxidative stress induced by Scopolamine. Biochemical estimation was performed to evaluate the changes in biochemical markers of Alzheimer’s disease such as lipid peroxidation (LPO), Glutathione reductase (GSH), and Catalase. The Scopolamine induced amnesia model has shown increased Acetylcholinesterase (AChE) levels and the inhibitory effect of test compounds in the brain AChE levels have been evaluated. In all the studies Donapezil (Dose: 50µg/kg) has been used as reference drug. The reduced AChE activity is shown by compounds 3f, 3c, and 3e. In the later stage, the most potent compounds have been evaluated for Aβ42 inhibitory profile. It can be hypothesized that this series of alkyl-aryl sulphonamides exhibit anti-AD activity by inhibition of Acetylcholinesterase (AChE) enzyme as well as inhibition of plaque formation on prolong dosage along with neuroprotection from oxidative stress.

Keywords: gamma-secretase inhibitors, Alzzheimer's disease, sulphonamides, QSAR

Procedia PDF Downloads 229

Authors: Asif Bilal, Fouzia Tanvir, Sibtain Ahmad

Abstract:

Estrogen receptor alpha, also known as estrogen receptor-1, is highly involved in risk of mammary carcinoma. The objectives of this study were to identify non-synonymous SNPs of estrogen receptor and their association with breast cancer and to identify the chemotherapeutic responses of phytochemicals against it via in-silico study design. For this purpose, different online tools. to identify pathogenic SNPs the tools were SIFT, Polyphen, Polyphen-2, fuNTRp, SNAP2, for finding disease associated SNPs the tools SNP&GO, PhD-SNP, PredictSNP, MAPP, SNAP, MetaSNP, PANTHER, and to check protein stability Mu-Pro, I-Mutant, and CONSURF were used. Post-translational modifications (PTMs) were detected by Musitedeep, Protein secondary structure by SOPMA, protein to protein interaction by STRING, molecular docking by PyRx. Seven SNPs having rsIDs (rs760766066, rs779180038, rs956399300, rs773683317, rs397509428, rs755020320, and rs1131692059) showing mutations on I229T, R243C, Y246H, P336R, Q375H, R394S, and R394H, respectively found to be completely deleterious. The PTMs found were 96 times Glycosylation; 30 times Ubiquitination, a single time Acetylation; and no Hydroxylation and Phosphorylation were found. The protein secondary structure consisted of Alpha helix (Hh) is (28%), Extended strand (Ee) is (21%), Beta turn (Tt) is 7.89% and Random coil (Cc) is (44.11%). Protein-protein interaction analysis revealed that it has strong interaction with Myeloperoxidase, Xanthine dehydrogenase, carboxylesterase 1, Glutathione S-transferase Mu 1, and with estrogen receptors. For molecular docking we used Asiaticoside, Ilekudinuside, Robustoflavone, Irinoticane, Withanolides, and 9-amin0-5 as ligands that extract from phytochemicals and docked with this protein. We found that there was great interaction (from -8.6 to -9.7) of these ligands of phytochemicals at ESR1 wild and two mutants (I229T and R394S). It is concluded that these SNPs found in ESR1 are involved in breast cancer and given phytochemicals are highly helpful against breast cancer as chemotherapeutic agents. Further in vitro and in vivo analysis should be performed to conduct these interactions.

Keywords: breast cancer, ESR1, phytochemicals, molecular docking

Procedia PDF Downloads 45

Authors: Vinod Kumar Gupta, Narendrakumar M. Chaudhari, Suchismitha Iskepalli, Chitra Dutta

Abstract:

Background-The community composition of the human microbiome is known to vary at distinct anatomical niches. But little is known about the nature of variations if any, at the genome/sub-genome levels of a specific microbial community across different niches. The present report aims to explore, as a case study, the variations in gene repertoire of 28 Prevotella reference draft genomes derived from different body-sites of human, as reported earlier by the Human Microbiome Consortium. Results-The analysis reveals the exclusive presence of 11798, 3673, 3348 and 934 gene families and exclusive absence of 17, 221, 115 and 645 gene families in Prevotella genomes derived from the human oral cavity, gastro-intestinal tracts (GIT), urogenital tract (UGT) and skin, respectively. The pan-genome for Prevotella remains “open”. Distribution of various functional COG categories differs appreciably among the habitat-specific genes, within Prevotella pan-genome and between the GIT-derived Bacteroides and Prevotella. The skin and GIT isolates of Prevotella are enriched in singletons involved in Signal transduction mechanisms, while the UGT and oral isolates show higher representation of the Defense mechanisms category. No niche-specific variations could be observed in the distribution of KEGG pathways. Conclusion-Prevotella may have developed distinct genetic strategies for adaptation to different anatomical habitats through selective, niche-specific acquisition and elimination of suitable gene-families. In addition, individual microorganisms tend to develop their own distinctive adaptive stratagems through large repertoires of singletons. Such in situ, habitat-driven refurbishment of the genetic makeup can impart substantial intra-lineage genome diversity within the microbes without perturbing their general taxonomic heritage.

Keywords: body niche adaptation, human microbiome, pangenome, Prevotella

Procedia PDF Downloads 231

Authors: Petre Voicu, Mircea Oaida, Radu Vasiu, Catalin Gheorghiu, Aurel Dumitru

Abstract:

The present work deals with the way in which food and tobacco waste can be used in agriculture. As a result of the lack of efficient technologies for their recycling, we are currently faced with the appearance of appreciable quantities of residual organic residues that find their use only very rarely and only after long storage in landfills. The main disadvantages of long storage of organic waste are the unpleasant smell, the high content of pathogenic agents, and the high content in the water. The release of these enormous amounts imperatively demands the finding of solutions to ensure the avoidance of environmental pollution. The measure practiced by us consists of the processing of this waste in special installations, testing in pilot experimental perimeters, and later administration on agricultural lands without harming the quality of the soil, agricultural crops, and the environment. The current crisis of raw materials and energy also raises special problems in the field of organic waste valorization, an activity that takes place with low energy consumption. At the same time, their composition recommends them as useful secondary sources in agriculture. The transformation of food scraps and other residues concentrated organics thus acquires a new orientation, in which these materials are seen as important secondary resources. The utilization of food and tobacco waste in agriculture is also stimulated by the increasing lack of chemical fertilizers and the continuous increase in their price, under the conditions that the soil requires increased amounts of fertilizers in order to obtain high, stable, and profitable production. The need to maintain and increase the humus content of the soil is also taken into account, as an essential factor of its fertility, as a source and reserve of nutrients and microelements, as an important factor in increasing the buffering capacity of the soil, and the more reserved use of chemical fertilizers, improving the structure and permeability for water with positive effects on the quality of agricultural works and preventing the excess and/or deficit of moisture in the soil.

Keywords: ecology, soil, organic waste, fertility

Procedia PDF Downloads 56

Authors: Rumaisa Shahid, Saad Aziz Durrani, Shameel Pervez, Ibatsam Khokhar

Abstract:

Food waste is a prevalent issue in Pakistan, with over 40 percent of food discarded annually. Despite their decay, rotting fruits retain residual nutritional value consumed by microorganisms, notably fungi and bacteria. Fungi, preferred for their extracellular enzyme release, are gaining prominence, particularly for pectinase production. This enzyme offers several advantages, including clarifying juices by breaking down pectic compounds. In this study, three Aspergillus flavus isolates derived from decomposed fruits and manure were selected for pectinase production. The primary aim was to isolate fungi from diverse waste sources, identify the isolates and assess their capacity for pectinase production. The identification was done through morphological characteristics with the help of Light microscopy and Scanning Electron Microscopy (SEM). Pectinolytic potential was screened using pectin minimal salt agar (PMSA) medium, comparing clear zone diameters among isolates. Identification relied on morphological characteristics. Optimizing substrate (lemon and orange peel powder) concentrations, pH, temperature, and incubation period aimed to enhance pectinase yield. Spectrophotometry enabled quantitative analysis. The temperature was set at room temperature (28 ºC). The optimal conditions for Aspergillus flavus strain AF1(isolated from mango) included a pH of 5, an incubation period of 120 hours, and substrate concentrations of 3.3% for orange peels and 6.6% for lemon peels. For AF2 and AF3 (both isolated from soil), the ideal pH and incubation period were the same as AF1 i.e. pH 5 and 120 hours. However, their optimized substrate concentrations varied, with AF2 showing maximum activity at 3.3% for orange peels and 6.6% for lemon peels, while AF3 exhibited its peak activity at 6.6% for orange peels and 8.3% for lemon peels. Among the isolates, AF1 demonstrated superior performance under these conditions, comparatively.

Keywords: pectinase, lemon peel, orange peel, aspergillus flavus

Procedia PDF Downloads 41

Authors: Sodamade Abiodun, Adeboye Olubunmi Omolara

Abstract:

Problems associated with protein malnutrition are still prevalent in third-world countries, leading to the constant search for plants that can serve as nutrients and medicinal purposes. Huckleberry is one of the plants that has been proven useful locally in the treatment of numerous ailments and diseases. A fresh sample of Huckleberry was collected from a vegetable garden situated near the Erelu dam of the Emmanuel Alayande College of Education campus, Oyo. The sample was authenticated at the forestry research institute of Nigeria (FRIN) Ibadan. The leaves of the plant were plucked and processed for leaf protein concentrates before proximate composition; mineral analysis phytochemical and antimicrobial properties of the leaf protein concentrates were determined using a standard method of analysis. The results of proximate constituents showed; moisture content; 9.89±0.051g/100g, Ash; 3.23±0.12g/100g, crude fat; 3.96±0.11g/100g and 61.27±0.56g/100g of Nitrogen free extractive results of the mineral analysis showed that the sample contains Mg; 0.081±0.00mg/100g, Ca; 42.30±0.05mg/100g, Na; 27.57±0.09mg/100g, K; 6.81±0.01mg/100g, P; 8.90±0.03mg/100g Fe; 0.51±0.00mg/100g, Zn; 0.021±0.00mg/100g, Cd; 0.04±0.04mg/100g, Pb; 0.002±0.00mg/100g, Cr; 0.041±0.00mg/100g while cadmium was not detected in the sample. The result of phytochemical analysis of leaf protein concentrates of the Huckleberry showed the presence of Alkaloid, Saponin, Flavonoid, Tanin, Coumarin, steroid, Terpenoid, cordial glycosides, Glycosides, Quinones, Anthocyanin, phytosterols, and phenols. Ethanolic extracts of the Huckleberry leaf protein concentrates showed that it contains bioactive compounds that are capable of eradicating some tested microorganisms; Staphylococcus aureus, Streptococcus pyogenes, Streptococcus faecalis, Pseudomonas aeruginosa, Klebisidlae pneumonia and Proteus merabilis. The results of the analysis of leaf protein concentrates of Huckleberry showed that the sample contains high nutrient and mineral constituents and phytochemical compounds that could make the sample useful for medicinal activities.

Keywords: huckleberry, mentha piperita, phytochemical, leaf protein concentrates, nutritional characteristics

Procedia PDF Downloads 61

Authors: Faiz Khan Mohammad, Saumya Ray Chaudhari, Raghunathan Rengaswamy, Swagatika Sahoo

Abstract:

Cholera caused by pathogenic gut bacteria Vibrio Cholerae (VC), is a major health problem in developing countries. Different strains of VC exhibit variable responses subject to different extracellular medium (Nag et al, Infect Immun, 2018). In this study, we present a new approach to model the variable VC responses in mono- and co-cultures, subject to continuously changing growth medium, which is otherwise difficult via simple FBA model. Nine VC strain and seven E. coli (EC) models were assembled and considered. A continuously changing medium is modelled using a new iterative-based controlled medium technique (ITC). The medium is appropriately prefixed with the VC model secretome. As the flux through the bacteria biomass increases secretes certain by-products. These products shall add-on to the medium, either deviating the nutrient potential or block certain metabolic components of the model, effectively forming a controlled feed-back loop. Different VC models were setup as monoculture of VC in glucose enriched medium, and in co-culture with VC strains and EC. Constrained to glucose enriched medium, (i) VC_Classical model resulted in higher flux through acidic secretome suggesting a pH change of the medium, leading to lowering of its biomass. This is in consonance with the literature reports. (ii) When compared for neutral secretome, flux through acetoin exchange was higher in VC_El tor than the classical models, suggesting El tor requires an acidic partner to lower its biomass. (iii) Seven of nine VC models predicted 3-methyl-2-Oxovaleric acid, mysirtic acid, folic acid, and acetate significantly affect corresponding biomass reactions. (iv) V. parhemolyticus and vulnificus were found to be phenotypically similar to VC Classical strain, across the nine VC strains. The work addresses the advantage of the ITC over regular flux balance analysis for modelling varying growth medium. Future expansion to co-cultures, potentiates the identification of novel interacting partners as effective cholera therapeutics.

Keywords: cholera, vibrio cholera El. tor, vibrio cholera classical, acetate

Procedia PDF Downloads 136

Authors: Ola A. Nassr, Ali M. Abd Alridha, Rua A. Naser, Rasha S. Abbas

Abstract:

Background: Excessive and inappropriate use of antimicrobial agents among hospitalized patients remains an important patient safety and public health issue worldwide. Not only does this behavior incur unnecessary cost but it is also associated with increased morbidity and mortality. The objective of this study is to obtain an insight into the prescribing patterns of antibiotics in surgical and medical wards, to help identify a scope for improvement in service delivery. Method: A simple point prevalence survey included a convenience sample of 200 patients admitted to medical and surgical wards in a government teaching hospital in Baghdad between October 2017 and April 2018. Data were collected by a trained pharmacy intern using a standardized form. Patient’s demographics and details of the prescribed antibiotics, including dose, frequency of dosing and route of administration, were reported. Patients were included if they had been admitted at least 24 hours before the survey. Patients under 18 years of age, having a diagnosis of cancer or shock, or being admitted to the intensive care unit, were excluded. Data were checked and entered by the authors into Excel and were subjected to frequency analysis, which was carried out on anonymized data to protect patient confidentiality. Results: Overall, 88.5% of patients (n=177) received 293 antibiotics during their hospital admission, with a small variation between wards (80%-97%). The average number of antibiotics prescribed per patient was 1.65, ranging from 1.3 for medical patients to 1.95 for surgical patients. Parenteral third-generation cephalosporins were the most commonly prescribed at a rate of 54.3% (n=159) followed by nitroimidazole 29.4% (n=86), quinolones 7.5% (n=22) and macrolides 4.4% (n=13), while carbapenems and aminoglycosides were the least prescribed together accounting for only 4.4% (n=13). The intravenous route was the most common route of administration, used for 96.6% of patients (n=171). Indications were reported in only 63.8% of cases. Culture to identify pathogenic organisms was employed in only 0.5% of cases. Conclusion: Broad-spectrum antibiotics are prescribed at an alarming rate. This practice may provoke antibiotic resistance and adversely affect the patient outcome. Implementation of an antibiotic stewardship program is warranted to enhance the efficacy, safety and cost-effectiveness of antimicrobial agents.

Keywords: Acute care, Antibiotic misuse, Iraq, Prescribing

Procedia PDF Downloads 103

Authors: Victor Ananth Paramananth, Rashid Muniginin, Mahaya Abd Rahman, Siti Afifah Ismail

Abstract:

Safety issues, scene protection, and appropriate evidence collection must be handled in any bio crime scene. There will be a scene or multi-scene to be cordoned for investigation in any bio-incident or bio crime event. Evidence collection is critical in determining the type of microbial or toxin, its lethality, and its source. As a consequence, from the start of the investigation, a proper sampling method is required. The most significant challenges for the crime scene officer would be deciding where to obtain samples, the best sampling method, and the sample sizes needed. Since there could be evidence in liquid, viscous, or powder shape at a crime scene, crime scene officers have difficulty determining which tools to use for sampling. To maximize sample collection, the appropriate tools for sampling methods are necessary. This study aims to assist the crime scene officer in collecting liquid, viscous, and powder biological samples in sufficient quantity while preserving sample quality. Observational tests on sample collection using liquid, viscous, and powder samples for adequate quantity and sample quality were performed using UV light in this research. The density of the light emission varies upon the method of collection and sample types. The best tools for collecting sufficient amounts of liquid, viscous, and powdered samples can be identified by observing UV light. Instead of active microorganisms, the invisible powder is used to assess sufficient sample collection during a crime scene investigation using various collection tools. The liquid, powdered and viscous samples collected using different tools were analyzed using Fourier transform infrared - attenuate total reflection (FTIR-ATR). FTIR spectroscopy is commonly used for rapid discrimination, classification, and identification of intact microbial cells. The liquid, viscous and powdered samples collected using various tools have been successfully observed using UV light. Furthermore, FTIR-ATR analysis showed that collected samples are sufficient in quantity while preserving their quality.

Keywords: biological sample, crime scene, collection tool, UV light, forensic

Procedia PDF Downloads 171

Authors: Gulnaz Khan, Meha F. Aftab, Munazza Murtaza, Rizwana S. Waraich

Abstract:

Advanced glycation end products (AGEs) precursor and its abnormal accumulation cause damage to various tissues and organs. AGEs have pathogenic implication in several diseases including diabetes. Existing AGEs inhibitors are not in clinical use, and there is a need for development of novel inhibitors. The present investigation aimed at identifying the novel AGEs inhibitors and assessing their mechanism of action for treating insulin resistance in mice model of diabetes. Novel derivatives of benzylidene of indan-1,3-dione were synthesized. The compounds were selected to study their action mechanism in improving insulin resistance, in vitro, in human hepatocytes and murine adipocytes and then, in vivo, in mice genetic model of diabetes (db/db). Mice were treated with novel derivatives of benzylidene of indane 1,3-dione. AGEs mediated ROS production was measured by dihydroethidium fluorescence assay. AGEs level in the serum of treated mice was observed by ELISA. Gene expression of receptor for AGEs (RAGE), PPAR-gamma, TNF-alpha and GLUT-4 was evaluated by RT-PCR. Glucose uptake was measured by fluorescent method. Microscopy was used to analyze glycogen synthesis in muscle. Among several derivatives of benzylidene of indan-1,3-dione, IDD-24, demonstrated highest inhibition of AGESs. IDD-24 significantly reduced AGEs formation and expression of receptor for advanced glycation end products (RAGE) in fat, liver of db/db mice. Suppression of AGEs mediated ROS production was also observed in hepatocytes and fat cell, after treatment with IDD-24. Glycogen synthesis was increased in muscle tissue of mice treated with IDD-24. In adipocytes, IDD-24 prevented AGEs induced reduced glucose uptake. Mice treated with IDD-24 exhibited increased glucose tolerance, serum adiponectin levels and decreased insulin resistance. The result of present study suggested that IDD-24 can be a possible treatment target to address glycotoxins induced insulin resistance.

Keywords: advance glycation end product, hyperglycemia, indan-1, 3-dione, insulin resistance

Procedia PDF Downloads 137

Authors: Geetika Vajpayee, Sugandha Asthana, Pratibha Kumari, Shanthy Sundaram

Abstract:

Pseudomonas species possess a variety of promising properties of antifungal and growth promoting activities in the wheat plant. In the present study, Pseudomonas fluorescens MTCC-9768 is tested against plant pathogenic fungus Tilletia indica, causing Karnal bunt, a quarantine disease of wheat (Triticum aestivum) affecting kernels of wheat. It is one of the 1/A1 harmful diseases of wheat worldwide under EU legislation. This disease develops in the growth phase by the spreading of microscopically small spores of the fungus (teliospores) being dispersed by the wind. The present chemical fungicidal treatments were reported to reduce teliospores germination, but its effect is questionable since T. indica can survive up to four years in the soil. The fungal growth inhibition tests were performed using Dual Culture Technique, and the results showed inhibition by 82.5%. The interaction of antagonist bacteria-fungus causes changes in the morphology of hyphae, which was observed using Lactophenol cotton blue staining and Scanning Electron Microscopy (SEM). The rounded and swollen ends, called ‘theca’ were observed in interacted fungus as compared to control fungus (without bacterial interaction). This bacterium was tested for its antagonistic activity like protease, cellulose, HCN production, Chitinase, etc. The growth promoting activities showed increase production of IAA in bacteria. The bacterial secondary metabolites were extracted in different solvents for testing its growth inhibiting properties. The characterization and purification of the antifungal compound were done by Thin Layer Chromatography, and Rf value was calculated (Rf value = 0.54) and compared to the standard antifungal compound, 2, 4 DAPG (Rf value = 0.54). Further, the in vivo experiments showed a significant decrease in the severity of disease in the wheat plant due to direct injection method and seed treatment. Our results indicate that the extracted and purified compound from the antagonist bacteria, P. fluorescens MTCC-9768 may be used as a potential biocontrol agent against T. indica. This also concludes that the PGPR properties of the bacteria may be utilized by incorporating it into bio-fertilizers.

Keywords: antagonism, Karnal bunt, PGPR, Pseudomonas fluorescens

Procedia PDF Downloads 371

Authors: Dawit Chankaye

Abstract:

Background: Azadirachta indica is the most versatile medicinal plant known as “the village pharmacy”. The plant is known for its broad spectrum of biological activity in India and various countries throughout history by many different human cultures. The present study was undertaken to determine the antimalarial and antidiabetic properties of the leaf extracts of A. indica grown in Ethiopia when treated in vivo. This work has also been concerned with determining essential oil composition and the antimicrobial activity of the plant in vitro. Methods: Leaf extracts were prepared using three different selected solvents. Standard and clinical isolates were treated with extracts of the leaves of A. indica using the agar well diffusion method. The antimalarial and antidiabetic tests were conducted in vivo in mice. Phytochemical screening was done using various chemical tests, and the volatile oil constituents were determined using gas chromatography-mass spectrometry (GC/MS). Results: In vivo antimalarial activity studies showed 85.23%, 69.01%, and 81.54% suppression of parasitemia for 70% ethanol, acetone, and water extracts, respectively. The extracts collected from the leaves also showed reduced blood sugar levels in alloxan-induced diabetic mice. In addition, the solvent extracts were shown to have an inhibitory effect on the growth of microorganisms under the study. The minimum inhibitory concentration (MIC) ranged from 850 to 1050 µg/ml. Notably, the phytochemical investigation of the ethanol extracts showed the presence of secondary metabolites. Seventeen compounds (mainly sesquiterpenes) that represent 75.45% of the essential oil were characterized by GC/MS analysis. Conclusion: Extracts examined in this study indicated that the leaf of A. indica grown in Ethiopia retained the biological activities demonstrating the extent equivalent to when it was grown in its natural habitat. In addition, phytochemical investigation and GC/MS analysis of volatile oil constituents showed comparable results to those presented in India and elsewhere.

Keywords: Azadirachta indica, vivo, antimalarial activity, antidiabetic activity, alloxan, mice, phytochemical

Procedia PDF Downloads 53

Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

Abstract:

Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

Procedia PDF Downloads 323

Authors: Asma Rashid, Shazia Iram, Iftikhar Ahmad

Abstract:

Mango sudden death is an emerging problem in Pakistan. As its prevalence is observed in almost all mango growing areas and severity varied from 2-5% in Punjab and 5-10% in Sindh. Symptoms on affected trees include bark splitting, discoloration of the vascular tissue, wilting, gummosis and at the end rapid death. Total of n= 45 isolates were isolated from different mango growing areas of Punjab and Sindh. Pathogenicity of these fungal isolates was tested through artificial inoculation method on different hosts (potato tubers, detached mango leaves, detached mango twigs and mango plants) under controlled conditions and all were proved pathogenic with varying degree of aggressiveness in reference to control. The findings of the present study proved that out of these four methods, potato tubers inoculation method was the most ideal as this fix the inoculums on the target site. Increased fungal growth and spore numbers may be due to soft tissues of potato tubers from which Ceratocystis isolates can easily pass. Lesion area on potato tubers was in the range of 7.09-0.14 cm2 followed by detached mango twigs which were ranged from 0.48-0.09 cm2). All pathological results were proved highly significant at P<0.05 through ANOVA but isolate to isolate showed non-significant behaviour but they have the positive effect on lesion area. Re-isolation of respective fungi was achieved with 100 percent success which results in the verification of Koch’s postulates. DNA of fungal pathogens was successfully extracted through phenol chloroform method. Amplification was done through ITS, b-tubulin gene, and Transcription Elongation Factor (EF1-a) gene primers and the amplified amplicons were sequenced and compared from NCBI which showed 99-100 % similarity with Ceratocystis manginecans. Fungus Ceratocystis manginecans formed one of strongly supported sub-clades through phylogenetic tree. Results obtained through this work would be supportive in establishment of relation of isolates with their region and will give information about pathogenicity level of isolates that would be useful to develop the management policies to reduce the afflictions in orchards caused by mango sudden death.

Keywords: artificial inoculation, mango, Ceratocystis manginecans, phylogenetic, screening

Procedia PDF Downloads 223

Authors: Raana Babadi Fathipour

Abstract:

The production of dairy holds significant importance in the sustenance of billions of individuals worldwide, as they rely on milk and its derived products for daily consumption. In addition to being a source of essential nutrients crucial for human well-being, such as proteins, fats, vitamins, and minerals; dairy items are witnessing an increasing demand worldwide. Amongst all the factors contributing to the quality and safety assurance of dairy products, the strong focus lies on maintaining high standards in raw milk procurement. Raw milk serves as an externally nutritious medium for various microorganisms due to its inherent properties. This poses a considerable challenge for the dairy industry in ensuring that microbial contamination is minimized throughout every stage of the value chain. Despite implementing diverse process technologies—both conventional and innovative—the occurrence of microbial spoilage still results in substantial losses within this industry context. Moreover, milk and dairy products have been associated with numerous cases of foodborne illnesses across the globe. Various pathogens such as Salmonella serovars, Campylobacter spp., Shiga toxin-producing Escherichia coli, Listeria monocytogenes, and enterotoxin producing Staphylococcus aureus are commonly identified as the culprits behind these outbreaks in the dairy industry. The effective management of food safety within this sector necessitates a proactive and risk-based approach to reform. However, this strategy presents difficulties for developing nations where informal value chains dominate the dairy sector. Whether operating on a small or large scale or falling within formal or informal realms, it is imperative that the dairy industry adheres to principles of good hygiene practices and good manufacturing practices. Additionally, identifying and managing potential sources of contamination is crucial in mitigating challenges pertaining to quality and safety precautions.

Keywords: dairy value chain, microbial contamination, food safety, hygiene

Procedia PDF Downloads 47

Authors: Lorenzo Cocola, Massimo Fedel, Dragiša Savić, Bojana Danilović, Luca Poletto

Abstract:

An instrument for the detection and evaluation of gaseous carbon dioxide in the headspace of closed containers has been developed in the context of Packsensor Italian-Serbian joint project. The device is based on Tunable Diode Laser Absorption Spectroscopy (TDLAS) with a Wavelength Modulation Spectroscopy (WMS) technique in order to accomplish a non-invasive measurement inside closed containers of fermented dairy products (yogurts and fermented cheese in cups and bottles). The purpose of this instrument is the continuous monitoring of carbon dioxide concentration during incubation and storage of products over a time span of the whole shelf life of the product, in the presence of different microorganisms. The instrument’s optical front end has been designed to be integrated in a thermally stabilized incubator. An embedded computer provides processing of spectral artifacts and storage of an arbitrary set of calibration data allowing a properly calibrated measurement on many samples (cups and bottles) of different shapes and sizes commonly found in the retail distribution. A calibration protocol has been developed in order to be able to calibrate the instrument on the field also on containers which are notoriously difficult to seal properly. This calibration protocol is described and evaluated against reference measurements obtained through an industry standard (sampling) carbon dioxide metering technique. Some sets of validation test measurements on different containers are reported. Two test recordings of carbon dioxide concentration evolution are shown as an example of instrument operation. The first demonstrates the ability to monitor a rapid yeast growth in a contaminated sample through the increase of headspace carbon dioxide. Another experiment shows the dissolution transient with a non-saturated liquid medium in presence of a carbon dioxide rich headspace atmosphere.

Keywords: TDLAS, carbon dioxide, cups, headspace, measurement

Procedia PDF Downloads 292

Authors: Aurora Gitto, Philipp Proksch

Abstract:

The presence of various microorganisms (bacteria, fungi) in surface water and groundwater represents an important issue for human health worldwide. The matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) has emerged as a promising and reliable tool for bacteria identification in clinical diagnostic microbiology and environmental strains thanks to an ionization technique that uses a laser energy absorbing matrix to create ions from large molecules with minimal fragmentation. The study aims first to conceptualise and set up library information and create a comprehensive database of MALDI-TOF-MS spectra from environmental water samples. The samples were analysed over a year (2021-2022) using membrane filtration methodology (0.45 μm and 0.22 μm) and then isolated on R2A agar for a period of 5 days and Yeast extract agar growing at 22 °C up to 4 days and 37 °C for 48 hours. The undetected organisms by MALDI-TOF-MS were analysed by PCR and then sequenced. The information obtained by the sequencing was further implemented in the MALDI-TOF-MS library. Among the culturable bacteria, the results show how the incubator temperature affects the growth of some genera instead of others, as demonstrated by Pseudomonas sp., which grows at 22 °C, compared to Bacillus sp., which is abundant at 37 °C. The bacteria community shows a variation in composition also between the media used, as demonstrated with R2A agar which has been defined by a higher presence of organisms not detected compared to YEA. Interesting is the variability of the Genus over one year of sampling and how the seasonality impacts the bacteria community; in fact, in some sampling locations, we observed how the composition changed, moving from winter to spring and summer. In conclusion, the bacteria community in groundwater and river bank filtration represents important information that needs to be added to the library to simplify future water quality analysis but mainly to prevent potential risks to human health.

Keywords: water quality, MALDI-TOF-MS, sequencing, library

Procedia PDF Downloads 57

Authors: Mudrika Singhal

Abstract:

The research paper is basically about how the genetically modified organisms evolved and their significance in today’s world. It also highlights about the various pros and cons of the genetically modified organisms and the progress of India in this field. A genetically modified organism is the one whose genetic material has been altered using genetic engineering techniques. They have a wide range of uses such as transgenic plants, genetically modified mammals such as mouse and also in insects and aquatic life. Their use is rooted back to the time around 12,000 B.C. when humans domesticated plants and animals. At that humans used genetically modified organisms produced by the procedure of selective breeding and not by genetic engineering techniques. Selective breeding is the procedure in which selective traits are bred in plants and animals and then are domesticated. Domestication of wild plants into a suitable cultigen is a well known example of this technique. GMOs have uses in varied fields ranging from biological and medical research, production of pharmaceutical drugs to agricultural fields. The first organisms to be genetically modified were the microbes because of their simpler genetics. At present the genetically modified protein insulin is used to treat diabetes. In the case of plants transgenic plants, genetically modified crops and cisgenic plants are the examples of genetic modification. In the case of mammals, transgenic animals such as mice, rats etc. serve various purposes such as researching human diseases, improvement in animal health etc. Now coming upon the pros and cons related to the genetically modified organisms, pros include crops with higher yield, less growth time and more predictable in comparison to traditional breeding. Cons include that they are dangerous to mammals such as rats, these products contain protein which would trigger allergic reactions. In India presently, group of GMOs include GM microorganisms, transgenic crops and animals. There are varied applications in the field of healthcare and agriculture. In the nutshell, the research paper is about the progress in the field of genetic modification, taking along the effects in today’s world.

Keywords: applications, mammals, transgenic, engineering and technology

Procedia PDF Downloads 575

Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

Abstract:

With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

Procedia PDF Downloads 47

Authors: Roberto Cassino, Valeria Dissette, Carlo Alberto Bignozzi, Daniele Pazzi

Abstract:

Background and Aims: The aim of this work was to investigate, both ‘in vitro’ and ‘in vivo’, if the new SCX technology (SiO2-Ag+Chlorex) can easily defeat infections and it is really more effective than SSD (SilverSulfaDiazine). ‘In vitro’ methods: we tested ‘in vitro’ the effectiveness of both silver materials using a pool of 5 strains: Pseudomonas Aeruginosa, Staphylococcus aureus, Escherichia Coli, Enterococcus hirae and Candida Albicans. 100 µl of this pool have been seeded on Petri dishes and kept for 24 hours in incubation at 37 C°. ‘In vivo’ methods: we enrolled patients with multiple infectious chronic wounds (according with cutting & harding criteria for infection); after a qualitative evaluation of the wounds bacterial population, taking a sample by plug, we included in the study 6 patients for a total of 10 wounds, infected by one or more of the microorganisms used for the ‘in vitro’ test. The protocol consisted of a treatment with a spray powder of SSD every 48 hours for 14 days; in case of worsening we should have to start a new treatment with a spray powder containing silicon dioxide, ionic silver and chlorexidine (SiO2-Ag+Chlorex) every 48 hours for 14 days. We evaluated the number of clinical signs of infection and the disappearance or not of the wound edge erithema. ‘In vitro’ results: SSD demonstrated a wide zone of inhibition within 24 hours, but after 5 days there was no more signs of inhibition; on the contrary SCX had a good inhibition ring that lasted more than 5 days. ‘In vivo’ results: all wounds treated with SSD got worse; the signs of infection increased and the wound edge erithema did not disappear. According with the protocol, we treated then all wounds with SCX and they all improved within the period of observation with complete disappearance of clinical signs of infection and no more wound edge erithema. Conclusions: the study demonstrated the effectiveness of SiO2-Ag+Chlorex, especially in terms of long lasting antimicrobial action. We had the same results ‘in vitro’, so that there has been a perfect correspondence between the laboratory outcomes and the clinical ones.

Keywords: chronic wounds, infections, ionic silver, SSD

Procedia PDF Downloads 295