Search results for: bacterial pathogens

Authors: Ilaisa Kacivakanadina, Samson Viulu, Brad Carte, Katy Soapi

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Two bioactive compounds namely Vulgamycin (also known as enterocin A) and 5-deoxyenterocin were purified from a marine bacterial strain 1903. Strain 1903 was isolated from marine sediments collected from the Solomon Islands. Morphological features of strain 1903 showed that it belongs to the genus Streptomyces. The two secondary metabolites were extracted using EtOAc and purified by chromatographic methods using EtOAc and hexane solvents. Mass spectrum and NMR data of pure compounds were used to elucidate the chemical structures. In this study, results showed that both compounds were strongly active against Wild Type Staphylococcus aureus (WTSA) (MIC < 1 µg/mL) and in Brine shrimp assays (BSA) (MIC < 1 µg/mL). 5-deoxyenterocin was also active against Rifamycin resistant Staphylococcus aureus (RRSA) (MIC, 250 µg/mL) while vulgamycin showed bioactivity against Methicillin resistant Staphylococcus aureus (MRSA) (MIC 250 µg/mL). To the best of our knowledge, this is the first study that showed the bio-activity of 5-deoxyenterocin. This is also the first time that Vulgamycin has been reported to be active in a BSA. There has not been any mechanism of action studies for these two compounds against pathogens. This warrants further studies on their mechanism of action against microbial pathogens.

Keywords: 5-deoxyenterocin, bioactivity, brine shrimp assay (BSA), vulgamycin

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Authors: Zhala Ahmad, Zainab Lazim, Haider Hamzah

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Bacterial resistance is a pressing global health issue, with multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) strains to pose a serious threat. In this context, researchers are investigating effective, safe, and affordable metabolites to combat these pathogens. This study focuses on gum essential oil (GEO) extracted from Pistacia atlantica and its activity and the mechanism of action against XDR Gram-negative Acinetobacter baumannii. GEO was extracted by hydrodistillation and analyzed using GC-MS. Eleven A. baumannii isolates were collected from the ward environment of Burn and Plastic Surgery Hospital in Al Sulaymaniyah City, Iraq. They were identified using the VITEK 2 system, 16S rRNA gene, and confirmed with the blaₒₓₐ₋₅₁ gene; A. baumannii ATCC 19606 was used as a reference strain. The isolates were identified as resistant to twelve different antibiotics spanning six distinct antibiotic classes while showing susceptibility to tetracycline and trimethoprim. Over 40 chemical constituents were detected in the gum's essential oils, with α-pinene being the most abundant. GEO was found to inhibit the growth of A. baumannii isolates; the minimum inhibitory concentration (MIC) of GEO was 2.5 µl/ml. GEO induced protein leakage, phosphate, and potassium ion efflux, distorted cell morphology, and cell death in the tested bacteria. GEO exhibited bacterial clearance and anti-adhesion activity using Band-Aids. This study's findings suggest that GEO could be used as a potential alternative treatment for infectious diseases caused by XRD pathogens, shedding further light on the importance of GEO in biomedical applications. Future studies must focus on generating clinically feasible sources of GEO for testing in small animal models before proceeding to human trials, ensuring safe and effective translation from the laboratory to the clinic.

Keywords: antibiotic resistance, Acinetobacter baumannii, essential oils, Pistacia atlantica, alpha-pinene

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Authors: Gopalu Karunakaran, Matheswaran Jagathambal, Nguyen Van Minh, Evgeny Kolesnikov, Denis Kuznetsov

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This work investigated the use of Beijerinckia indica extracellular extract for the synthesis of silver nanoparticles using AgNO₃. The formation of nanoparticles was confirmed by different methods, such as UV-Vis absorption spectroscopy, XRD, FTIR, EDX, and TEM analysis. The formation of silver nanoparticles (AgNPs) was confirmed by the change in color from light yellow to dark brown. The absorbance peak obtained at 430 nm confirmed the presence of silver nanoparticles. The XRD analysis showed the cubic crystalline phase of the synthesized nanoparticles. FTIR revealed the presence of groups that acts as stabilizing and reducing agents for silver nanoparticles formation. The synthesized silver nanoparticles were generally found to be spherical in shape with size ranging from 5 to 20 nm, as evident by TEM analysis. These nanoparticles were found to inhibit pathogenic bacterial strains. This work proved that the bacterial extract is a potential eco-friendly candidate for the synthesis of silver nanoparticles with promising antibacterial and antioxidant properties. 

Keywords: antioxidant activity, antimicrobial activity, Beijerinckia indica, characterisation, extracellular extracts, silver nanoparticles

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Authors: G. Petruzzelli, F. Pedron, M. Grifoni, A. Pera, I. Rosellini, B. Pezzarossa

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The addition of lime as Ca(OH)2 to sewage sludge to destroy pathogens (Escherichia coli), was evaluated also in relation to heavy metal bioavailability. The obtained results show that the use of calcium hydroxide at the dose of 3% effectively destroyed pathogens ensuring the stability at high pH values over long period and the duration of the sewage sludge stabilization. In general, lime addition decreased the total extractability of heavy metals indicating a reduced bioavailability of these elements. This is particularly important for a safe utilization in agricultural soils to reduce the possible transfer of heavy metals to the food chain.

Keywords: biological sludge, Ca(OH)2, copper, pathogens, sanitation, zinc

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Authors: Shigenori Togashi, Kei Takenaka

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We developed new processes which can collect and detect rapidly airborne pathogens such as the avian flu virus for the pandemic prevention. The fluorescence antibody technique is known as one of high-sensitive detection methods for viruses, but this needs up to a few hours to bind sufficient fluorescence dyes to viruses for detection. In this paper, we developed a mist-labeling can detect substitution viruses in a short time to improve the binding rate of fluorescent dyes and substitution viruses by the micro reaction process. Moreover, we developed the rapid detection system with the above 'mist labeling'. The detection system set with a sampling bag collecting patient’s breath and a cartridge can detect automatically pathogens within 10 minutes.

Keywords: viruses, sampler, mist, detection, fluorescent dyes, microreaction

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Authors: Shimayali Kaushal, Nitesh Priyadarshi, Nitin Kumar Singhal

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Food borne bacterial species have been identified as major pathogens in most of the severe pathogen-related diseases among humans which result in great loss to human health and food industry. Conventional methods like plating and enzyme-linked immune sorbent assay (ELISA) are time-consuming, laborious and require specialized instruments. Nanotechnology has emerged as a great field in case of rapid detection of pathogens in recent years. The AuNRs material has good electro-optical properties due to its larger light absorption band and scattering in surface plasmon resonance wavelength regions. By exploiting the sugar-based adhesion properties of microorganism, we can use the glycoconjugates capped gold nanorods as a potential nanobiosensor to detect the foodborne pathogen. In the present study, polyethylene glycol (PEG) coated gold nanorods (AuNRs) were prepared and functionalized with different types of carbohydrates and further characterized by UV-Visible spectrophotometry, dynamic light scattering (DLS), transmission electron microscopy (TEM). The reactivity of above said nano-biosensor was probed by lectin binding assay and also by different strains of foodborne bacteria by using spectrophotometric and microscopic techniques. Due to the specific interaction of probe with foodborne bacteria (Escherichia coli, Pseudomonas aeruginosa), our nanoprobe has shown significant and selective ablation of targeted bacteria. Our findings suggest that our nanoprobe can be an ideal candidate for selective optical detection of food pathogens and can reduce loss to the food industry.

Keywords: glyco-conjugates, gold nanorods, nanobiosensor, nanoprobe

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

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

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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Authors: Geeta Negi, Pankaj, Anjana Srivastava, Anita Sharma

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In the present study, the presence of endosulfan, imidacloprid, carbendazim, in the soil /vegetables/cereals and water samples was observed in agriculture fields of Uttarakhand. In view of biodegradation of these pesticides, nine bacterial isolates were recovered from the soil samples of the fields which tolerated endosulfan, imidacloprid, carbendazim from 100 to 200 µg/ml. Three bacterial consortia used for in vitro bioremediation experiments were three bacterial isolates for carbendazim, imidacloprid and endosulfan, respectively. Maximum degradation (87 and 83%) of α and β endosulfan respectively was observed in soil slurry by consortium. Degradation of Imidacloprid and carbendazim under similar conditions was 88.4 and 77.5% respectively. FT-IR analysis of biodegraded samples of pesticides in liquid media showed stretching of various bonds. GC-MS of biodegraded endosulfan sample in soil slurry showed the presence of non-toxic intermediates. A pot trial with Bacterial treatments lowered down the uptake of pesticides in onion plants.

Keywords: biodegradation, carbendazim, consortium, endosulfan

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Authors: J. Y. Ijato, A. Adewumi, H. O Yakubu, O. O. Olajide, B. O. Ojo, B. A. Adanikin

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Post-harvest fruit rot pathogens are one of the major factors that are responsible for food security challenges in developing countries like Nigeria. These pathogens also cause fruit food poisoning. Biocidal effects of ethanolic extracts of Khaya grandifoliola, Hyptis suaveolens, Zingiber officinale, Calophyllum inophyllum, Datura stramonium on the mycelia growth of fungal rot pathogens of pineapple fruit was investigated, the ethanolic extracts of these test plants exhibited high significant inhibitory effects on the rot pathogens, the highest ethanolic extract inhibition of Zingiber officinale was on Aspergillus flavus (38.40%) at 1.0g/ml while the least inhibitory effect was on Aspergillus fumigatus (23.10%) at 1.0g/ml, the highest ethanol extract inhibition of Datura stramonium was on Aspergillus tubingensis (24.00%) at 1.0g/ml while the least inhibitory effect was 10.00% on Colletotrichum fruticola at 1.0g/ml, the highest ethanol extract inhibition of Calophyllum inophyllum was on Trichoderma harzianum (18.50%) at 1.0g/ml while the least inhibitory effect was on Aspergillus flavus (15.00%) at 1.0g/ml, the highest ethanol extract inhibition of Hyptis suaveolens was on Aspergillus fumigatus (35.00%) at 1.0g/ml while the least inhibitory effect was on Aspergillus niger (20.00%) at 1.0g/ml, the highest ethanol extract inhibition of Khaya grandifoliola was on Aspergillus flavus (35.00%) at 1.00g/ml while the least inhibitory effect was on Aspergillus fumigates (22.00%) at 1.0g/ml, the antifungal capacity of these test plant extracts on rot causing fungi on pineapple fruit reveals the possibility of their use by farmers and fruit traders as alternative to chemical fungicide that portends great threat to human and environmental health.

Keywords: fruit rot, pathogens, plant extracts, pineapple, food poisoning

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Authors: Soheir Mohamed Fathey

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Biofilms are complex biological communities which are hard to be eliminated by conventional antibiotic administration and implemented in eighty percent of humans infections. Green remedies have been used for centuries and have shown obvious effects in hindering and combating microbial biofilm infections. Nowadays, there has been a growth in the number of researches on the anti-biofilm performance of natural agents such as plant essential oil (EOs) and propolis. In this study, we investigated the antibiofilm performance of various natural agents, including four essential oils (EOs), cinnamon (Cinnamomum cassia), tea tree (Melaleuca alternifolia), and clove (Syzygium aromaticum), as well as propolis versus the biofilm of both Gram-positive pathogenic bacterium Staphylococcus aureus and Gram-negative pathogenic bacterium Pseudomonas aeruginosa which are major human and animal pathogens rendering a high risk due to their biofilm development ability. The antibiofilm activity of the tested agents was evaluated by crystal violet staining assay and detected by scanning electron and fluorescent microscopy. Antibiofilm performance declared a potent effect of the tested products versus the tested bacterial biofilms.

Keywords: biofilm, essential oils, electron microscopy, fluorescent

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Authors: Gimhani Madhushika Hewayalage, Thilini Ariyadasa, Sanja Gunawardena

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In Sri Lanka, the largest contribution for the industrial export earnings is governed by textile and apparel industry. However, this industry generates huge quantities of effluent consists of unfixed dyes which enhance the effluent colour and toxicity thereby leading towards environmental pollution. Therefore, the effluent should properly be treated prior to the release into the environment. The biological technique has now captured much attention as an environmental-friendly and cost-competitive effluent decolourization method due to the drawbacks of physical and chemical treatment techniques. The present study has focused on identifying dye decolourizing potential of several bacterial isolates obtained from the effluent of the local textile industry. Yellow EXF, Red EXF, Blue EXF, Nova Black WNN and Nylosan-Rhodamine-EB dyes have been selected for the study to represent different chromophore groups such as Azo and Xanthene. The rates of decolorization of each dye have been investigated by employing distinct bacterial isolates. Bacterial isolate which exhibited effective dye decolorizing potential was identified as Proteus mirabilis using 16S rRNA gene sequencing analysis. The high decolorizing rates of identified bacterial strain indicate its potential applicability in the treatment of dye-containing wastewaters.

Keywords: azo, bacterial, biological, decolourization, xanthene

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Authors: Vivek Bhat, Rohini Kelkar, Sanjay Biswas

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Introduction: Cancer patients are at increased risk of bacterial infections. This may due to the disease process itself, the effect of chemotherapeutic drugs or invasive procedures such as catheterization. A wide variety of bacteria including some emerging pathogens are increasingly being reported from these patients. The incidence of multidrug-resistant organisms particularly in the Gram negative group is also increasing, with higher resistance rates seen to cephalosporins, β-lactam/β-lactam inhibitor combinations, and the carbapenems. This study documents the bacteriological spectrum of infections and their resistance patterns in cancer patients. Methods: This study includes all bacterial isolates recovered from infections cancer patients over a period of 18 months. Samples included Blood cultures, Pus/wound swabs, urine, tissue biopsies, body fluids, catheter tips and respiratory specimens such as sputum and bronchoalveolar lavage (BAL). All samples were processed in the microbiology laboratory as per standard laboratory protocols. Organisms were identified to species level and antimicrobial susceptibility testing was performed manually by the disc diffusion technique or in the Vitek-2 (Biomereux, France) instrument. Interpretations were as per Clinical laboratory Standards Institute (CLSI) guidelines. Results: A total of 1150 bacterial isolates were cultured from 884 test samples during the study period. Of these 227 were Gram-positive and 923 were Gram-negative organisms. Staphylococcus aureus (99 isolates) was the commonest Gram-positive isolate followed by Enterococcus (79) and Gr A Streptococcus (30). Among the Gram negatives, E. coli (304), Pseudomonas aeruginosa (201) and Klebsiella pneumoniae (190) were the most common. Of the Staphylococcus aureus isolates 27.2% were methicillin resistant. Only 5.06% enterococci were vancomycin resistant. High rates of resistance to cefotaxime and ciprofloxacin were seen amongst E. coli (84.8% & 83.55%) and Klebsiella pneumoniae (71 & 62.1%) respectively. Resistance to carbapenems (meropenem) was high at 70% in Acinetobacter spp.; however all isolates were sensitive to colistin. Among the aminoglycosides, amikacin retained good efficacy against Escherichia coli (82.9%) and Pseudomonas aeruginosa (78.1%). Occasional isolates of emerging pathogens such as Chryseobacterium indologens, Roseomonas, and Achromobacter xyloxidans were also recovered. Conclusion: The common infections in cancer patients include respiratory, wound, tract infections and sepsis. The commonest isolates include Staphylococcus aureus, Enterococci, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. There is a high level of resistance to the commonly used antibiotics among Gram-negative organisms.

Keywords: bacteria, resistance, infection, cancer

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Authors: Sanaa Bardaweel

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Inorganic polyphosphate (poly P) is present in all living forms tested to date, from each of the three kingdoms of life. Studied mainly in prokaryotes, poly P and its associated enzymes are vital in diverse basic metabolism, in at least some structural functions and, notably, in stress responses. These plentiful and unrelated roles for poly P are probably the consequence of its presence in life-forms early in evolution. The genomes of many bacterial species, including pathogens, encode a homologue of a major poly P synthetic enzyme, poly P kinase 1 (PPK1). Genetic deletion of ppk1 results in reduced poly P levels and loss of pathogens virulence towards protozoa and animals. Thus far, no PPK1 homologue has been identified in higher-order eukaryotes and, therefore, PPK1 represents a novel target for chemotherapy. The idea of the current study is to purify the PPK1 from Escherichia coli to homogeneity in order to study the effect of active site point mutations on PPK1 catalysis via the application of site-directed mutagenesis strategy. The knowledge obtained about the active site of PPK1 will be utilized to characterize the catalytic and kinetic mechanism of PPK1 with model substrates. Comprehensive understanding of the enzyme kinetic mechanism and catalysis will be used to design and screen a library of synthetic compounds for potential discovery of selective PPK1-inhibitors.

Keywords: antimicobial, Escherichia coli, inorganic polyphosphate, PPK1-inhibitors

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Authors: Sofia Papadimitriou

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INTRODUCTION: The differential diagnosis between acute viral and bacterial infections is an important cost-effectiveness parameter at the stage of the treatment process in order to achieve the maximum benefits in therapeutic intervention by combining the minimum cost to ensure the proper use of antibiotics.The discovery of sensitive and robust molecular diagnostic tests in response to the role of the host in infections has enhanced the accurate diagnosis and differentiation of infections. METHOD: The study used a sample of six independent blood samples (total=756) which are associated with human proteins-proteins, each of which at the transcription stage expresses a different response in the host network between viral and bacterial infections.Τhe individual blood samples are subjected to a sequence of computer filters that identify a gene panel corresponding to an autonomous diagnostic score. The data set and the correspondence of the gene panel to the diagnostic patents a new Bangalore -Viral Bacterial (BL-VB). FINDING: We use a biomarker based on the blood of 10 genes(Panel-VB) that are an important prognostic value for the detection of viruses from bacterial infections with a weighted average AUROC of 0.97(95% CL:0.96-0.99) in eleven independent samples (sets n=898). We discovered a base with a patient score (VB 10 ) according to the table, which is a significant diagnostic value with a weighted average of AUROC 0.94(95% CL: 0.91-0.98) in 2996 patient samples from 56 public sets of data from 19 different countries. We also studied VB 10 in a new cohort of South India (BL-VB,n=56) and found 97% accuracy in confirmed cases of viral and bacterial infections. We found that VB 10 (a)accurately identifies the type of infection even in unspecified cases negative to the culture (b) shows its clinical condition recovery and (c) applies to all age groups, covering a wide range of acute bacterial and viral infectious, including non-specific pathogens. We applied our VB 10 rating to publicly available COVID 19 data and found that our rating diagnosed viral infection in patient samples. RESULTS: Τhe results of the study showed the diagnostic power of the biomarker VB 10 as a diagnostic test for the accurate diagnosis of acute infections in recovery conditions. We look forward to helping you make clinical decisions about prescribing antibiotics and integrating them into your policies management of antibiotic stewardship efforts. CONCLUSIONS: Overall, we are developing a new property of the RNA-based biomarker and a new blood test to differentiate between viral and bacterial infections to assist a physician in designing the optimal treatment regimen to contribute to the proper use of antibiotics and reduce the burden on antimicrobial resistance, AMR.

Keywords: acute infections, antimicrobial resistance, biomarker, blood transcriptome, systems biology, classifier diagnostic score

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Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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Authors: Muhammad Younis Laghari

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Global climate change has had many effects on the aquatic environment, and the major issue is pollution. Along with the other pollutants, there are a significant number of human microbial pathogens that pollute the water bodies. Another concern about the water quality is that the major aquatic resources bring water-borne pathogens and other related diseases. These resources include industrial effluent, untreated domestic sewage, acid mine drainage, etc. However, these water discharges through various routes may have treatment to eliminate the pathogenic microbes. Therefore, it is essential to control the leakage from sewer systems, residential discharge, and agricultural run-off. These pathogenic microbes have been implicated in the lives of water health (fishes), which is harmful and causes diseases. Mostly, the mortality of aquatic species results because of catastrophic floods due to poor water waste treatment and sanitation that introduce pathogenic bacteria into rivers. Pathogens survive in rivers and remain poorly known but essential to control water-borne diseases. The presence of bacteria in watercourses is diverse and constitutes a complicated subject. Many species are autochthonous and play an important role in aquatic ecosystems, while many others arise from untreated or poorly treated waste from industrial and domestic sources. Further, more investigation is required to know the induction of water-borne pathogens in various water resources and the potential impacts of water resource development on pathogen contamination.

Keywords: microbial pathogens, contamination, water resources, river water body

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Authors: Rupa Rani, Vipin Kumar

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Endosulfan, an organochlorine pesticide is of environmental concern due to its apparent persistence and toxicity. It has been reported as contaminants in soil, air, and water and is bioaccumulated and magnified in ecosystems. The combined use of microorganisms and plants has great potential for remediating soil contaminated with organic compounds such as pesticides. The objective of this study was to evaluate whether the bacterial inoculation influences plant growth promotion, endosulfan degradation in soil and endosulfan accumulation in different plant parts. Lycopersicon esculentum L. (Tomato) was grown in endosulfan spiked soil and inoculated with endosulfan tolerant bacterial strains. Endosulfan residues from different parts of plants and soil were extracted and estimated by using gas chromatograph equipped with 63Ni electron capture detector (GC-ECD). The inoculation of bacterial strains into the soil with plants showed a beneficial effect on endosulfan degradation and plant biomass production. Maximum endosulfan (90%) degradation was observed after 120 days of bacterial inoculation in the soil. Furthermore, there was significantly less endosulfan accumulation in roots and shoots of bacterial strains inoculated plants as compared to uninoculated plants. The results show the effectiveness of inoculated endosulfan tolerant bacterial strains to increase the remediation of endosulfan contaminated soil.

Keywords: organochlorine pesticides, endosulfan, degradation, plant-bacteria partnerships

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Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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Authors: S. Techaoei, K. Jarmkom, P. Eakwaropas, W. Khobjai

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The objective of this research was focused on investigating in vitro antimicrobial activity of Phellinus linteus fruiting body extracts on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Phellinus linteus fruiting body was extracted with ethanol and ethyl acetate and was vaporized. The disc diffusion assay was used to assess antimicrobial activity against tested bacterial strains. Primary screening of chemical profile of crude extract was determined by using thin layer chromatography. The positive control and the negative control were used as erythromycin and dimethyl sulfoxide, respectively. Initial screening of Phellinus linteus crude extract with the disc diffusion assay demonstrated that only ethanol had greater antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The MIC assay showed that the lower MIC was observed with 0.5 mg/ml of Pseudomonas aeruginosa and Methicillin-resistant Staphylococcus aureus and 0.25 mg/ml. of Escherichia coli and Staphylococcus aureus, respectively. TLC chemical profile of extract was represented at R_f ≈ 0.71-0.76.

Keywords: Staphylococcus aureus, Escherichia coli, Phellinus linteus, Methicillin-resistant Staphylococcus aureus, antimicrobial activity

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Authors: Mehani Mouna, Ladjel segni

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Humans use plants for thousands of years to treat various ailments, In many developing countries, Much of the population relies on traditional doctors and their collections of medicinal plants to cure them. Essential oils have many therapeutic properties. In herbal medicine, They are used for their antiseptic properties against infectious diseases of fungal origin, Against dermatophytes, Those of bacterial origin. The aim of our study is to determine the antimicrobial effect of essential oils of the plant Schinus molle on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, They are used for their antiseptic properties against infectious diseases of fungal origin, Against dermatophytes, Those of bacterial origin. The test adopted is based on the diffusion method on solid medium (Antibiogram), This method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plant Schinus molle has a different effect on the resistance of germs: For Pseudomonas aeruginosa strain is a moderately sensitive with an inhibition zone of 10 mm, Further Antirobactere, Escherichia coli and Proteus are strains that represent a high sensitivity, A zone of inhibition equal to 14.66 mm.

Keywords: Essential oil, microorganism, antibiogram, shinus molle

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Authors: Zikora K. G. Anyaegbunam, Annabel A. Nnawuihe, Ngozi J. Anyaegbunam, Emmanuel A. Eze

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The discovery and production of new antibiotics is unavoidable in the fight against drug-resistant bacteria. However, this is only part of the problem; we have never really had medications that could completely eradicate an infection. All pathogens create a limited number of dormant persister cells that are resistant to antibiotic treatment. When the concentration of antibiotics decreases, surviving persisters repopulate the population, resulting in a recurrent chronic infection. Bacterial populations have an alternative survival strategy to withstand harsh conditions or antibiotic exposure, in addition to the well-known methods of antibiotic resistance and biofilm formation. Persister cells are a limited subset of transiently antibiotic-tolerant phenotypic variations capable of surviving high-dose antibiotic therapy. Persisters that flip back to a normal phenotype can restart growth when antibiotic pressure drops, assuring the bacterial population's survival. Persister cells have been found in every major pathogen, and they play a role in antibiotic tolerance in biofilms as well as the recalcitrance of chronic infections. Persister cells has been implicated to play a role in the establishment of antibiotic resistance, according to growing research. Thusthe need to basically elucidate the biology of persisters and how they are linked to antibiotic resistance, and as well it's link to diseases.

Keywords: persister cells, phenotypic variations, repopulation, mobile genetic transfers, antibiotic resistance

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Authors: S. Susanti, V. P. Bintoro, A. Setiadi, S. I. Santoso, D. R. Febriandi

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Meat is a foodstuff of animal origin. It is perishable because a suitable medium for bacterial growth. That is why meat can be a potential hazard to humans. Several ways have been done to inhibit bacterial population in an effort to prolong the meat shelf-life. However, aberration sometimes happens in the practices of meat preservation, for example by using chemical material that possessed strong antibacterial activity like formaldehyde. For health reason, utilization of formaldehyde as a food preservative was forbidden because of DNA damage resulting cancer and birth defects. Therefore, it is important to seek a natural food preservative that is not harmful to the body. This study aims to reveal the potency of cashew apple as natural food preservative by measuring its antibacterial activity and marinating effect on the bacterial growth of beef and chicken meat. Antibacterial activity was measured by The Kirby-Bauer method while bacterial growth was determined by total plate count method. The results showed that inhibition zone of 10-30% cashew apple extract significantly wider compared to 0% extract on the medium of E. coli, S. aureus, S. typii, and Bacillus sp. Furthermore, beef marinated with 20-30% cashew apple extract and chicken meat marinated with 5-15% extract significantly less in the total number of bacteria compared to 0% extract. It can be concluded that marinating with 5-30% cashew apple extract can effectively inhibit the bacterial growth of beef and chicken meat. Moreover, the concentration of extracts to inhibit bacterial populations in chicken meat was reached at the lower level compared to beef. Thus, cashew apple is potential as a natural food preservative.

Keywords: bacterial growth, cashew apple, marinating, meat

Procedia PDF Downloads 239

Authors: Azila Adnan, Giridhar R. Nair, Mark C. Lay, Janis E. Swan

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The effect of aeration on bacterial cellulose (BC) production by Gluconacetobacter xylinus DSM46604 was studied in 5-L bioreactor. Four aeration rates were applied (0.3, 0.6, 1.0 and 1.5 vvm) in the fermentation media at constant agitation rate, 150 rpm. One vvm enhanced BC concentration (4.4 g/L) and productivity (0.44 g/L/day) while greater agitation rate (1.5 vvm) decreased BC concentration (4.0 g/L) and productivity (0.40 g/L/day).

Keywords: aeration, bacterial cellulose, fermentation, gluconacetobacter xylinus

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Authors: Elizabeth Starzycka-Korbas, Michal Starzycki, Wojciech Rybinski, Mirosława Dabert

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For a few years, the populations of pathogenic fungi occurring in winter oilseed rape in Malyszyn were analyzed. Brassica napus L. in Poland and in the world is a source of energy for both the men (oil), and animals, as post-extraction middling, as well as a motor fuel (oil, biofuel) therefore studies of this type are very important. The species composition of pathogenic fungi can be an indicator of seed yield. The occurrence of oilseed rape pathogens during several years were analyzed using the sequencing method DNA ITS. The results were compared in the gene bank using the program NCBI / BLAST. In field conditions before harvest of oilseed rape presence of pathogens infesting B. napus has been assessed. For example, in 2015, 150 samples have been isolated and applied to PDA medium for the identification of belonging species. From all population has been selected mycelium of 83 isolates which were sequenced. Others (67 isolates) were pathogenic fungi of the genus Alternaria which are easily to recognize. The population of pathogenic species on oilseed rape have been identified after analyzing the DNA ITS and include: Leptosphaeria sp. 38 (L. maculans 25, L. biglobosa 13), Alternaria sp. 29, Fusarium sp. 3, Sclerotinia sclerotiorum 7, heterogeneous 6, total of 83 isolates. The genus Alternaria sp. fungi wear the largest share of B. napus pathogens in particular years. Another dangerous species for oilseed rape was Leptosphaeria sp. Populations of pathogens in each year were different. The number of pathogens occurring in the field and their composition is very important for breeders and farmers because of the possible selection of the most resistant genotypes for sowing in the next growing season.

Keywords: B. napus, DNA ITS Sequencing, pathogenic fungi, population

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Authors: Alloysius C. Ogodo, Ositadinma C. Ugbogu, Uzochukwu G. Ekeleme

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Food poisoning and infection by bacteria are of public health significance to both developing and developed countries. Samples of ogi (akamu) prepared from white and yellow variety of maize sold in Uturu and Okigwe were analyzed together with the laboratory prepared ogi for microbial quality using the standard microbiological methods. The analyses showed that both white and yellow variety had total bacterial counts (cfu/g) of 4.0 ×107 and 3.9 x 107 for the laboratory prepared ogi while the commercial ogi had 5.2 x 107 and 4.9 x107, 4.9 x107 and 4.5 x107, 5.4 x107 and 5.0 x107 for Eke-Okigwe, Up-gate and Nkwo-Achara market respectively. The Staphylococcal counts ranged from 2.0 x 102 to 5.0 x102 and 1.0 x 102 to 4.0 x102 for the white and yellow variety from the different markets while Staphylococcal growth was not recorded on the laboratory prepared ogi. The laboratory prepared ogi had no Coliform growth while the commercially prepared ogi had counts of 0.5 x103 to 1.6 x 103 for white variety and 0.3 x 103 to 1.1 x103 for yellow variety respectively. The Lactic acid bacterial count of 3.5x106 and 3.0x106 was recorded for the laboratory ogi while the commercially prepared ogi ranged from 3.2x106 to 4.2x106 (white variety) and 3.0 x106 to 3.9 x106 (yellow). The presence of bacteria isolates from the commercial and laboratory fermented ogi showed that Lactobacillus sp, Leuconostoc sp and Citrobacter sp were present in all the samples, Micrococcus sp and Klebsiella sp were isolated from Eke-Okigwe and ABSU-up-gate markets varieties respectively, E. coli and Staphylococcus sp were present in Eke-Okigwe and Nkwo-Achara markets while Salmonella sp were isolated from the three markets. Hence, there are chances of contracting food borne diseases from commercially prepared ogi. Therefore, there is the need for sanitary measures in the production of fermented cereals so as to minimize the rate of food borne pathogens during processing and storage.

Keywords: ogi, fermentation, bacterial quality, lactic acid bacteria, maize

Procedia PDF Downloads 382

Authors: Michelle S. Pereira, Analice C. Azevedo, Julliane D. Medeiros, Ana Claudia S. Martins, Didier S. Castellano-Filho, Claudio G. Diniz, Vania L. Silva

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Vaginal microbiota is a complex ecosystem, composed by aerobic and anaerobic bacteria, living in a dynamic equilibrium. Lactobacillus spp. are predominant in vaginal ecosystem, and factors such as immunity and hormonal variations may lead to disruptions, resulting in proliferation of opportunistic pathogens. Bacterial vaginosis (BV) is a polymicrobial syndrome, caused by an increasing of anaerobic bacteria replacing Lactobacillus spp. Microorganisms such as Gardnerella vaginalis, Mycoplasma hominis, Mobiluncus spp., and Atopobium vaginae can be found in BV, which may also be associated to other infections such as by Human Papillomavirus (HPV). HPV is highly prevalent in sexually active women, and is considered a risk factor for development of cervical cancer. As long as few data is available on vaginal microbiota of women with HPV-associated cervical lesions, our objectives were to evaluate the diversity in vaginal ecosystem in these women. To all patients, clinical and socio-demographic data were collected after gynecological examination. This study was approved by the Ethics Committee from Federal University of Juiz de Fora, Minas Gerais, Brazil. Vaginal secretion and cervical scraping were collected. Gram-stained smears were evaluated to establish Nugent score for BV determination. Viral and bacterial DNA obtained was used as template for HPV genotyping (PCR) and bacterial fingerprint (REP-PCR). In total 31 patients were included (mean age 35 and 93.6% sexually active). The Nugent score showed that 38.7% were BV. From the medical records, Pap smear tests showed that 32.3% had low grade squamous epithelial lesion (LSIL), 29% had high grade squamous epithelial lesion (HSIL), 25.8% had atypical squamous cells of undetermined significance (ASC-US) and 12.9% with atypical squamous cells that would not exclude high-grade lesion (ASC-H). All participants were HPV+. HPV-16 was the most frequent (87.1%), followed by HPV-18 (61.3%). HPV-31, HPV-52 and HPV-58 were also detected. Coinfection HPV-16/HPV-18 was observed in 75%. In the 18-30 age group, HPV-16 was detected in 40%, and HPV-16/HPV-18 coinfection in 35%. HPV-16 was associated to 30% of ASC-H and 20% of HSIL patients. BV was observed in 50% of HPV-16+ participants and in 45% of HPV-16/HPV-18+. Fingerprints of bacterial communities showed clusters with low similarity suggesting high heterogeneity in vaginal microbiota within the sampled group. Overall, the data is worrisome once cervical-cancer highly risk-associated HPV-types were identified. The high microbial diversity observed may be related to the different levels of cellular lesions, and different physiological conditions of the participants (age, social behavior, education). Further prospective studies are needed to better address correlations and BV and microbial imbalance in vaginal ecosystems which would be related to the different cellular lesions in women with HPV infections. Supported by FAPEMIG, CNPq, CAPES, PPGCBIO/UFJF.

Keywords: human papillomavirus, bacterial vaginosis, bacterial diversity, cervical cancer

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Authors: Yogesh Karunakar, Praveen Kandaswamy

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Mother’s milk provides the most superior source of nutrition to a child. There is no other substitute to the mother’s milk. Postpartum secretions like breast milk can be analyzed on the go for testing the presence of any harmful pathogen before a mother can feed the child or donate the milk for the milk bank. Since breast feeding is one of the main causes for transmission of diseases to the newborn, it is mandatory to test the secretions. In this paper, we describe the detection of pathogens like E-coli, Human Immunodeficiency Virus (HIV), Hepatitis B (HBV), Hepatitis C (HCV), Cytomegalovirus (CMV), Zika and Ebola virus through an innovative method, in which we are developing a unique chip for testing the mother’s milk sample. The chip will contain an antibody specific to the target pathogen that will show a color change if there are enough pathogens present in the fluid that will be considered dangerous. A smart-phone camera will then be acquiring the image of the strip and using various image processing techniques we will detect the color development due to antigen antibody interaction within 5 minutes, thereby not adding to any delay, before the newborn is fed or prior to the collection of the milk for the milk bank. If the target pathogen comes positive through this method, then the health care provider can provide adequate treatment to bring down the number of pathogens. This will reduce the postpartum related mortality and morbidity which arises due to feeding infectious breast milk to own child.

Keywords: postpartum, fluids, camera, HIV, HCV, CMV, Zika, Ebola, smart-phones, breast milk, pathogens, image processing techniques

Procedia PDF Downloads 200

Authors: Khaled Khleifat

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This study detects the bacterial species that responsible for UTI in both diabetic patients and non-diabetic patients, Jordan. 116 urine samples were investigated in order to determine UTI-causing bacteria. These samples distributed unequally between diabetic male (12) and diabetic female (25) and also non-diabetic male (13) and non-diabetic female (66). The results represent that E.coli is responsible for UTI in both diabetic and non-diabetic patients (15.5% and 29.3% respectively) with large proportion (44.8%). This study showed that not all bacterial species that isolated from the non-diabetic sample could be isolated from diabetic samples. E. coli (15.5%), P. aeruginosa (4.3%), K. pneumonia (1.7%), P. mirabilis (2.6%), S. marcescens (0.9%), S. aureus (1.7%), S. pyogenes (1.7%), E. faecalis (0.9%), S. epidermidis (1.7%) and S. saprophyticus (0.9%). But E. aerogenes, E. cloacae, C. freundii, A. baumannii and B. subtilis are five bacterial species that can’t isolate from all diabetic samples. This study shows that for the treatment of UTI in both diabetic and non-diabetic patients, Chloramphenicol (30 μg), Ciprofloxacin (5 μg) and Vancomycin (30 μg) are more favorable than other antibiotics. In the same time, Cephalothin (30μg) is not recommended.

Keywords: urinary tract infections, diabetes mellitus, bacterial species, infections

Procedia PDF Downloads 301

Authors: Amel Bouderhem, Aminata Ould El Hadj Khelil, Amina N. Djrarbaoui, Aroussi Aroussi

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The present work aims to find the influence of the nature of the soil on the effectiveness of the biodegradation of hydrocarbons by a mixture of bacterial strains hydrocarbonoclastes. Processes of bioaugmentation and biostimulation trial are applied to samples of soils polluted voluntarily by the crude oil. For the evaluation of the biodegradation of hydrocarbons, the bacterial load, the pH and organic carbon total are followed in the different experimental batches. He bacterial load of the sandy soil varies among the witnesses of 45,2 .108 CFU/ml at the beginning of the experimentation to 214,07.108 CFU/ml at the end of the experiment. Of the soil silty-clay varies between 103,31 .108 CFU/ml and 614,86.108 CFU/ml . It was found a strong increase in the bacterial biomass during the processing of all samples. This increase is more important in the samples of sand bioaugmente or biomass increased from 63.16 .108 CFU/ml to 309.68 .108 CFU/ml than in soil samples silty clay- bioaugmente whose content in bacteria evolved of 73,01 .108 CFU/ml to 631.80 . 108CFU/ml

Keywords: pollution, hydrocarbons, bioremediation, bacteria hydrocarbonoclastes, ground, texture

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Authors: Li Cheng, Zhang Yilei, Cohen Yehuda

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Two motility mechanisms were introduced into iDynoMiCs software, which adopts an individual-based modeling method. Based on the new capabilities, along with the pressure motility developed before, influence of bacterial motility on biofilm formation was studied. Simulation results were evaluated both qualitatively through 3D structure inspections and quantitatively by parameter characterizations. It was showed that twitching motility increased the biofilm surface irregularity probably due to movement of cells towards higher nutrient concentration location whereas free motility, on the other hand, could make biofilms flatter and smoother relatively. Pressure motility showed no significant influence in this study.

Keywords: iDynoMics, biofilm structure, bacterial motility, motility mechanisms

Procedia PDF Downloads 356