Search results for: resistant microbial strains

Authors: Michael Kvopka, Ezekiel Kingston

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A 61-year-old man with no significant past medical history presented to a quaternary ophthalmic referral center with acute right-sided medial canthal pain, periorbital edema, and erythema despite oral antibiotic therapy. CT imaging confirmed the presence of right preseptal cellulitis and lacrimal sac aspiration identified multi-resistant Enterobacter cloacae. A diagnosis of acute right-sided dacryocystitis with preseptal cellulitis was made. He was successfully treated with broadening of antibiotic therapy to intravenous meropenem. The symptomatic resolution was noted on follow-up without evidence of disease recurrence. To the Authors’ best knowledge, this is the first reported case of multi-resistant E. cloacae dacryocystitis and preseptal cellulitis. The management of this patient required a multi-disciplinary approach, so the Authors believe this report is relevant to general ophthalmologists and oculoplastic sub-specialists.

Keywords: enterobacter, dacryocystitis, preseptal cellulitis, antibiotic resistance

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Authors: Saria Almarzook, Monika Reissmann, Gudrun Brockmann

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Due to its history, occurrence in different ecosystems and diverse using, the modern horse (Equus caballus) shows large variability in size, appearance, behavior and habits. At all times, breeders try to create groups (breeds, strains) representing high homology but showing clear differences in comparison to other groups. A great interest of analyzing phenotypic and genetic traits looking for real diversity and genetic uniqueness existents for Arabian horses in Syria. 90 Arabian horses from governmental research center of Arabian horses in Damascus were included. The horses represent three strains (Kahlawi, Saklawi, Hamdani) originated from different geographical zones. They were raised on the same farm, under stable conditions. Twelve phenotypic traits were measured: wither height (WH), croup width (CW), croup height (CH), neck girth (NG), thorax girth (TG), chest girth (ChG), chest depth (ChD), chest width (ChW), back line length (BLL), body length (BL), fore cannon length (FCL) and hind cannon length (HCL). The horses were divided into groups according to age (less than 2 years, 2-4 years, 4-9 years, over 9 years) and to sex (male, female). The statistical analyzes show that age has significant influence of WH while the strain has only a very limited effect. On CW, NG, BLL, FCL and HCL, there is only a significant influence of sex. Age has significant effect on CH and BL. All sources of classes have a significant effect on TG, ChG, ChD and ChW. Strain has a significant effect on the BL. These results provide first information for real biodiversity in and between the strains and can be used to develop the breeding work in the Arabian horse breed.

Keywords: Arabian horse, phenotypic traits, strains, Syria

Procedia PDF Downloads 363

Authors: K. Karapetyan, F. Tkhruni, A. Aghajanyan, T. S. Balabekyan, L. Arstamyan

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Introduction: Globalization of the food supply has created the conditions favorable for the emergence and spread of food-borne and especially dangerous pathogens (EDP) in developing countries. The fresh-cut fruit and vegetable industry is searching for alternatives to replace chemical treatments with biopreservative approaches that ensure the safety of the processed foods product. Antimicrobial compounds of lactic acid bacteria (LAB) possess bactericidal or bacteriostatic activity against intestinal pathogens, spoilage organisms and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus and Salmonella. Endemic strains of LAB were isolated. The strains, showing broad spectrum of antimicrobial activity against food spoiling microorganisms, were selected. The genotyping by 16S rRNA sequencing, GS-PCR, RAPD PCR methods showed that they were presented by Lactobacillus rhamnosus109, L.plantarum 65, L.plantarum 66 and Enterococcus faecium 64 species. LAB are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB strains were isolated from different dairy products from rural households from the highland regions of Armenia. Serially diluted samples were spread on MRS (Merck, Germany) and hydrolyzed milk agar (1,2 % w/v). Single colonies from each LAB were individually inoculated in liquid MRS medium and incubated at 37oC for 24 hours. Culture broth with biomass was centrifuged at 10,000 g during 20 min for obtaining of cell free culture broth (CFC). The antimicrobial substances from CFC broth were purified by the combination of adsorption-desorption and ion-exchange chromatography methods. Separation of bacteriocins was performed using a HPLC method on "Avex ODS" C18 column. Mass analysis of peptides recorded on the device API 4000 in the electron ionization mode. The spot-on-lawn method on the test culture plated in the solid medium was applied. The antimicrobial activity is expressed in arbitrary units (AU/ml). Results. Purification of CFC broth of LAB allowed to obtain partially purified antimicrobial preparations which contains bacteriocins with broad spectrum of antimicrobial activity. Investigation of their main biochemical properties shown, that inhibitory activity of preparations is partially reduced after treatment with proteinase K, trypsin, pepsin, suggesting a proteinaceous nature of bacteriocin-like substances containing in CFC broth. Preparations preserved their activity after heat treatment (50-121 oC, 20 min) and were stable in the pH range 3–8. The results of SDS PAAG electrophoresis show that L.plantarum 66 and Ent.faecium 64 strains have one bacteriocin (BCN) with maximal antimicrobial activity with approximate molecular weight 2.0-3.0 kDa. From L.rhamnosus 109 two BCNs were obtained. Mass spectral analysis indicates that these bacteriocins have peptide bonds and molecular weight of BCN 1 and BCN 2 are approximately 1.5 kDa and 700 Da. Discussion: Thus, our experimental data shown, that isolated endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms of different taxonomic group, such as Salmonella sp., Esherichia coli, Bacillus sp., L.monocytogenes, Proteus mirabilis, Staph. aureus, Ps. aeruginosa. Obtained results proved the perspectives for use of endemic strains in the preservation of foodstuffs. Acknowledgments: This work was realized with financial support of the Project Global Initiatives for Preliferation Prevention (GIPP) T2- 298, ISTC A-1866.

Keywords: antimicrobial activity, bacteriocins, endemic strains, food safety

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Authors: I. Savitskaya, A. Kistaubayeva, A. Zhubanova, I. Blavachinskaiya, D. Ibrayeva, M. Abdulzhanova, A. Otarbay, A.Isabekova

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This study was conducted for the investigation of number of cellulolytic bacteria and their ability in decomposition. Seven samples surface soil were collected on cellulose Zailiskii Alatau slopes. Cellulolitic activity of new strains of Bacillus, isolated from soil is determined. Isolated cellulose degrading bacteria were screened for determination of the highest cellulose activity by quantitative assay using Congo red, gravimetric assay and colorimetric DNS method trough of the determination of the parameters of sugar reduction. Strains are assigned to: B.subtilis, B.licheniformis, B. cereus and, В. megaterium. Bacillus strains consisting of several different types of cellulases have broad substrate specificity of cellulase complexes formed by them. Cellulolitic bacteria were recorded to have highest cellulase activity and selected for optimization of cellulase enzyme production.

Keywords: cellulose-degrading bacteria, cellulase complex, foothills soil, screening

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Authors: Ji-Sun Lee, Soong-Keun Hyun, Jin-Ho Kim

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In this study, a glass fiber is fabricated using a continuous spinning process from alkali resistant (AR) glass with 4 wt% zirconia. In order to confirm the melting properties of the marble glass, the raw material is placed into a Pt crucible and melted at 1650 ℃ for 2 h, and then annealed. In order to confirm the transparency of the clear marble glass, the visible transmittance is measured, and the fiber spinning condition is investigated by using high temperature viscosity measurements. A change in the diameter is observed according to the winding speed in the range of 100–900 rpm; it is also verified as a function of the fiberizing temperature in the range of 1200–1260 ℃. The optimum winding speed and spinning temperature are 500 rpm and 1240 ℃, respectively. The properties of the prepared spinning fiber are confirmed using optical microscope, tensile strength, modulus, and alkali-resistant tests.

Keywords: glass composition, fiber diameter, continuous filament fiber, continuous spinning, physical properties

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Authors: Sihame Amakrane, Zineb Ouahdi, Mohammed Salah, Said Belaaouad

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\This research explores the efficacy of novel pyrimidine derivatives on bacterial strains such as Escherichia coli, Staphylococcus aureus, and Myccobacterium tuberculosis, utilizing bending energy calculations. Of the 25 compounds examined, 13 displayed potent activity against all the bacterial strains under study, exhibiting bending energy measurements between -7.4 and -10.7 kcal/mol. The -7.4 kcal/mol value corresponds to the bending energy of the SA12 and SA13 compounds with the 2xct protein (Staphylococcus aureus), whereas the -10.7 kcal/molis linked with the bending energy of SA6 and SA11 compounds with the 6GAV protein (Myccobacterium tuberculosis). Further research will involve a QSAR (Quantitative Structure-Activity Relationship) study aimed at constructing a reliable model to combat the aforementioned bacterial strains and a molecular dynamics study to evaluate the stability of ligand-protein complexes.

Keywords: docking, QSAR, bending energy, e. coli

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Authors: Lok Bahadur Shrestha, Narayan Raj Bhattarai, Basudha Khanal

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Introduction: Coagulase-negative staphylococci (CoNS) are the normal commensal of human skin and mucous membranes. The study was carried out to study the prevalence of CoNS among clinical isolates, to characterize them up to species level and to compare the three conventional methods for detection of biofilm formation. Objectives: to characterize the clinically significant coagulase-negative staphylococci up to species level, to compare the three phenotypic methods for the detection of biofilm formation and to study the antimicrobial susceptibility pattern of the isolates. Methods: CoNS isolates were obtained from various clinical samples during the period of 1 year. Characterization up to species level was done using biochemical test and study of biofilm formation was done by tube adherence, congo red agar, and tissue culture plate method. Results: Among 71 CoNS isolates, seven species were identified. S. epidermidis was the most common species followed by S. saprophyticus, S. haemolyticus. Antimicrobial susceptibility pattern of CoNS documented resistance of 90% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 55% of the isolates. We detected biofilm formation in 71.8% of isolates. The sensitivity of tube adherence method was 82% while that of congo red agar method was 78%. Conclusion: Among 71 CoNS isolated, S. epidermidis was the most common isolates followed by S. saprophyticus and S. haemolyticus. Biofilm formation was detected in 71.8% of the isolates. All of the methods were effective at detecting biofilm-producing CoNS strains. Biofilm former strains are more resistant to antibiotics as compared to biofilm non-formers.

Keywords: CoNS, congo red agar, bloodstream infections, foreign body-related infections, tissue culture plate

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Authors: Sajjad Jafari

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Background and Aim: The use of herbal medicines has a long history. Today, due to the resistance of microorganisms to antibiotics and antimicrobial substances, herbal medicines have attracted attention due to their significant antimicrobial effects and low toxicity. This study aims to systematically review the antimicrobial effects of different plant extracts (Quercus infectoria) as possible candidates for treating infectious diseases. Material and Methods: The present study is a review study by searching reputable scientific databases such as PubMed, Google Scholar, Scopus, and Web of Science from 2000 to 2023 using the keywords Antimicrobial, Quercus infectoria, Medicinal herbal, Infectious diseases the latest information obtained. Results: In this study, 45 articles were found and reviewed. Quercus infectoria is a small tree native to Greece, Asia Minor, and Iran. Quercus is a plant genus in the family of Fagaceae. This species is generally known under the name ‘‘baloot” in Iran and is commonly used as a medicinal plant. The extracts used included water, hydro-alcoholic, ethanol, methanol. This plant had high inhibition activity and a lethal effect on gram-positive and gram-negative bacteria of ATCC strains, hospital, and resistant strains. Therefore, in addition to antibacterial effects, antiparasitic and antifungal effects. The seed of the plant was the most used and the most effective antimicrobial extract among the ethanol and methanol extracts. Conclusion: The findings of this study suggest that Quercus infectoria has significant antimicrobial effects against a wide range of microorganisms. This makes it a potential candidate for the development of new antimicrobial drugs. Further research is needed to confirm the efficacy and safety of Quercus infectoria in clinical trials.

Keywords: antimicrobial, Quercus infectoria, medicinal herbal, infectious diseases

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Authors: M. Doyeni, S. Suproniene, V. Tilvikiene

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Agricultural wastes contribute significantly to global climate change through greenhouse gas emissions if not adequately recycled and sustainably managed. A recurring agricultural waste is livestock wastes that have consistently served as feedstock for biogas systems. The objective of this study was to access the influence of digestate fertilization on soil microbial activity and greenhouse gas emissions in agricultural fields. Wheat (Triticum spp. L.) was fertilized with different types of animal wastes digestates (organic fertilizers) and mineral nitrogen (inorganic fertilizer) for three years. The 170 kg N ha⁻¹ presented in digestates were split fertilized at an application rate of 90 and 80 kg N ha⁻¹. The soil microorganism activity could be predicted significantly using the dehydrogenase activity and soil microbial biomass carbon. By combining the two different monitoring approaches, the different methods applied in this study were sensitive to enzymatic activities and organic carbon in the living component of the soil organic matter. The emissions of greenhouse gasses (carbon dioxide (CO₂), methane (CH₄), and nitrous oxide (N₂O) were monitored directly by a static chamber system. The soil and environmental variables were measured to determine their influence on greenhouse gas emissions. Emission peaks was observed in N₂O and CO₂ after the first application of fertilizers with the emissions flattening out over the cultivating season while CH₄ emission was negligible with no apparent patterns observed. Microbial biomass carbon and dehydrogenase activity were affected by the fertilized organic digestates. A significant difference was recorded between the control and the digestate treated soils for the microbial biomass carbon and dehydrogenase. Results also showed individual and cumulative emissions of CO₂, CH₄ and N₂O from the digestates were relatively low suggesting the digestate fertilization can be an efficient method for improving soil quality and reducing greenhouse gases from agricultural sources in temperate climate conditions.

Keywords: greenhouse gas emission, manure digestate, soil microbial activity, yield

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Authors: Vagolu S. Krishna, Shan Zheng, Estharla M. Rekha, Luke W. Guddat, Dharmarajan Sriram

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Tuberculosis (TB) remains a major threat to human health. This due to the fact that current drug treatments are less than optimal as well as the rising occurrence of multi drug-resistant and extensively drug-resistant strains of the etiological agent, Mycobacterium tuberculosis (Mt). Given the wide-spread significance of this disease, we have undertaken a design and evaluation program to discover new anti-TB drug leads. Here, our attention is focused on ketol-acid reductoisomerase (KARI), the second enzyme in the branched-chain amino acid biosynthesis pathway. Importantly, this enzyme is present in bacteria but not in humans, making it an attractive proposition for drug discovery. In the present work, we used high-throughput virtual screening to identify seventeen potential inhibitors of KARI using the Birla Institute of Technology and Science in-house database. Compounds were selected based on high docking scores, which were assigned as the result of favourable interactions between the compound and the active site of KARI. The Ki values for two leads, compounds 14 and 16 are 3.71 and 3.06 µM, respectively for Mt KARI. To assess the mode of binding, 100 ns molecular dynamics simulations for these two compounds in association with Mt KARI were performed and showed that the complex was stable with an average RMSD of less than 2.5 Å for all atoms. Compound 16 showed an MIC of 2.06 ± 0.91 µM and a 1.9 fold logarithmic reduction in the growth of Mt in an infected macrophage model. The two compounds exhibited low toxicity against murine macrophage RAW 264.7 cell lines. Thus, both compounds are promising candidates for development as an anti-TB drug leads.

Keywords: ketol-acid reductoisomerase, macrophage, molecular docking and dynamics, tuberculosis

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Authors: Syahidah N. Zulkifli, Herlina A. Rahim, Nurul A. M. Subha

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Sustaining water quality is critical in order to avoid any harmful health consequences for end-user consumers. The detection of microbial impurities at the household level is the foundation of water security. Water quality is now monitored only at water utilities or infrastructure, such as water treatment facilities or reservoirs. This research provides a first-hand scientific understanding of microbial composition presence in Malaysia’s household point-of-use (POUs) water supply influenced by seasonal fluctuations, standstill periods, and flow dynamics by using the NIR-Raman spectroscopic technique. According to the findings, 20% of water samples were contaminated by pathogenic bacteria, which are Legionella and Salmonella cells. A comparison of the spectra reveals significant signature peaks (420 cm⁻¹ to 1800 cm⁻¹), including species-specific bands. This demonstrates the importance of regularly monitoring POUs water quality to provide a safe and clean water supply to homeowners. Conventional Raman spectroscopy, up-to-date, is no longer suited for real-time monitoring. Therefore, this study introduced an alternative micro-spectrometer to give a rapid and sustainable way of monitoring POUs water quality. Assessing microbiological threats in water supply becomes more reliable and efficient by leveraging IoT protocol.

Keywords: microbial contaminants, water quality, water monitoring, Raman spectroscopy

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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae bacteria is a global threat to human health due to an increase in multi-drug resistance among strains. The hypervirulent strains of Klebsiella pneumoniae is a major trouble due to their association with life-threatening infections in a healthy population. One of the major virulence factors of hyper virulent strains of Klebsiella pneumoniae is the T6SS (Type six secretary system) which is majorly involved in microbial antagonism and causes interaction with the host eukaryotic cells during infections. T6SS mediates some of the crucial factors for establishing infection by the bacteria, such as cell adherence, invasion, and subsequent in vivo colonisation. The antibacterial activity and the cell invasion property of the T6SS system is a major requirement for the establishment of K. pneumoniae infections within the gut. The T6SS can be an appropriate target for developing therapeutics. The T6SS consists of an inner tube comprising hexamers of Hcp (Haemolysin -regulated protein) protein, and at the top of this tube sits VgrG (Valine glycine repeat protein G); the tip of the machinery consists of PAAR domain containing proteins which act as a delivery system for bacterial effectors. For this study, immune response to recombinant VgrG protein was generated to establish this protein as a potential immunogen for the development of therapeutic leads. The immunogenicity of the selected protein was determined by predicting the B cell epitopes by the BCEP analysis tool. The gene sequence for multiple domains of VgrG protein (phage_base_V, T6SS_Vgr, DUF2345) was selected and cloned in pMAL vector in E. coli. The construct was subcloned and expressed as a fusion protein of 203 residue protein with mannose binding protein tag (MBP) to enhance solubility and purification of this protein. The purified recombinant VgrG fusion protein was used for mice immunisation. The antiserum showed reactivity with the recombinant VgrG in ELISA and western blot. The immunised mice were challenged with K. pneumoniae bacteria and showed bacterial clearance in immunised mice. The recombinant VgrG protein can further be used for studying downstream signalling of VgrG protein in mice during infection and for therapeutic MAb development to eradicate K. pneumoniae infections.

Keywords: immune response, Klebsiella pneumoniae, multi-drug resistance, recombinant protein expression, T6SS, VgrG

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Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

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Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

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Authors: V. Ivanov, J. Chu, V. Stabnikov

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Low cost, sustainable, and environmentally friendly microbial cements, grouts, polysaccharides and bioplastics are useful in construction and geotechnical engineering. Construction-related biotechnologies are based on activity of different microorganisms: urease-producing, acidogenic, halophilic, alkaliphilic, denitrifying, iron- and sulphate-reducing bacteria, cyanobacteria, algae, microscopic fungi. The bio-related materials and processes can be used for the bioaggregation, soil biogrouting and bioclogging, biocementation, biodesaturation of water-satured soil, bioencapsulation of soft clay, biocoating, and biorepair of the concrete surface. Altogether with the most popular calcium- and urea based biocementation, there are possible and often are more effective such methods of ground improvement as calcium- and magnesium based biocementation, calcium phosphate strengthening of soil, calcium bicarbonate biocementation, and iron- or polysaccharide based bioclogging. The construction-related microbial biotechnologies have a lot of advantages over conventional construction materials and processes.

Keywords: ground improvement, biocementation, biogrouting, microorganisms

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Authors: Anshu Siwach, Qianlai Zhuang, Ratul Baishya

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Context: This study focuses on the influence of moss cover and seasonality on soil microbial biomass and enzymatic activity in different Central Himalayan temperate forest types. Soil microbial biomass and enzymes are key indicators of microbial communities in soil and provide information on soil properties, microbial status, and organic matter dynamics. The activity of microorganisms in the soil varies depending on the vegetation type and environmental conditions. Therefore, this study aims to assess the effects of moss cover, seasons, and different forest types on soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN), and soil enzymatic activity in the Central Himalayas, Uttarakhand, India. Research Aim: The aim of this study is to evaluate the levels of SMBC, SMBN, and soil enzymatic activity in different temperate forest types under the influence of two ground covers (soil with and without moss cover) during the rainy and winter seasons. Question Addressed: This study addresses the following questions: 1. How does the presence of moss cover and seasonality affect soil microbial biomass and enzymatic activity? 2. What is the influence of different forest types on SMBC, SMBN, and enzymatic activity? Methodology: Soil samples were collected from different forest types during the rainy and winter seasons. The study utilizes the chloroform-fumigation extraction method to determine SMBC and SMBN. Standard methodologies are followed to measure enzymatic activities, including dehydrogenase, acid phosphatase, aryl sulfatase, β-glucosidase, phenol oxidase, and urease. Findings: The study reveals significant variations in SMBC, SMBN, and enzymatic activity under different ground covers, within the rainy and winter seasons, and among the forest types. Moss cover positively influences SMBC and enzymatic activity during the rainy season, while soil without moss cover shows higher values during the winter season. Quercus-dominated forests, as well as Cupressus torulosa forests, exhibit higher levels of SMBC and enzymatic activity, while Pinus roxburghii forests show lower levels. Theoretical Importance: The findings highlight the importance of considering mosses in forest management plans to improve soil microbial diversity, enzymatic activity, soil quality, and health. Additionally, this research contributes to understanding the role of lower plants, such as mosses, in influencing ecosystem dynamics. Conclusion: The study concludes that moss cover during the rainy season significantly influences soil microbial biomass and enzymatic activity. Quercus and Cupressus torulosa dominated forests demonstrate higher levels of SMBC and enzymatic activity, indicating the importance of these forest types in sustaining soil microbial diversity and soil health. Including mosses in forest management plans can improve soil quality and overall ecosystem dynamics.

Keywords: moss cover, seasons, soil enzymes, soil microbial biomass, temperate forest types

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Authors: Azna Zuberi

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Biofilm is a sessile bacterial accretion in which bacteria adapts different physiological and morphological behavior from planktonic form. It is the root cause of about 80% microbial infections in human. Among them, E. coli biofilms are most prevalent in medical devices associated nosocomial infections. The objective of this study was to inhibit biofilm formation by targeting LuxS gene, involved in quorum sensing using CRISPRi. luxS is a synthase, involved in the synthesis of Autoinducer-2(AI-2), which in turn guides the initial stage of biofilm formation. To implement CRISPRi system, we have synthesized complementary sgRNA to target gene sequence and co-expressed with dCas9. Suppression of luxS was confirmed through qRT-PCR. The effect of luxS gene on biofilm inhibition was studied through crystal violet assay, XTT reduction assay and scanning electron microscopy. We conclude that CRISPRi system could be a potential strategy to inhibit bacterial biofilm through mechanism base approach.

Keywords: biofilm, CRISPRi, luxS, microbial

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Authors: Surajbhan Sevda

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Metal containing wastewater is generated in large quintiles due to rapid industrialization. Generally, the metal present in wastewater is not biodegradable and can be accumulated in living animals, humans and plant tissue, causing disorder and diseases. The conventional metal recovery methods include chemical, physical and biological methods, but these are chemical and energy intensive. The recent development in microbial fuel cell (MFC) technology provides a new approach for metal recovery; this technology offers a flexible platform for both reduction and oxidation reaction oriented process. The use of MFCs will be a new platform for more efficient and low energy approach for metal recovery from the wastewater. So far metal recover was extensively studied using chemical, physical and biological methods. The MFCs present a new and efficient approach for removing and recovering metals from different wastewater, suggesting the use of different electrode for metal recovery can be a new efficient and effective approach.

Keywords: metal recovery, microbial fuel cell, wastewater, bioelectricity

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Authors: Shashank Gahlaut, Chandrashekhar Sharan, J. P. Singh

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Increasing incidence of antibiotic-resistant bacteria is a big concern for the treatment of pathogenic diseases. The effect of treatment of patients with antibiotics often leads to the evolution of antibiotic resistance in the pathogens. The detection of antibiotic or antimicrobial resistant bacteria (microbes) is quite essential as it is becoming one of the big threats globally. Here we propose a novel technique to tackle this problem. We are taking a step forward to prevent the infections and diseases due to drug resistant microbes. This detection is based on some unique features of silver (a noble metal) nanorods (AgNRs) which are fabricated by a physical deposition method called thermal glancing angle deposition (GLAD). Silver nanorods are found to be highly sensitive and selective for hydrogen sulfide (H2S) gas. Color and water wetting (contact angle) of AgNRs are two parameters what are effected in the presence of this gas. H₂S is one of the major gaseous products evolved in the bacterial metabolic process. It is also known as gasotransmitter that transmits some biological singles in living systems. Nitric Oxide (NO) and Carbon mono oxide (CO) are two another members of this family. Orlowski (1895) observed the emission of H₂S by the bacteria for the first time. Most of the microorganism produce these gases. Here we are focusing on H₂S gas evolution to determine live/dead and antibiotic-resistant bacteria. AgNRs array has been used for the detection of H₂S from micro-organisms. A mobile app is also developed to make it easy, portable, user-friendly, and cost-effective.

Keywords: antibiotic resistance, hydrogen sulfide, live and dead bacteria, mobile app

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Authors: S. N. Al-Bahry, S. K. Al-Musharafi, I. Y. Mahmoud

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Recycled treated sewage effluents (TSE) is used for agriculture, Public park irrigation and industrial purposes. TSE was found to play a major role in the distribution of antibiotic resistant bacteria into the environment. Fecal coliform and enterococci counts were significantly higher during summer compared to winter seasons. Oman has low annual rainfall with annual average temperature varied between 15-45oC. The main source of potable water is from seawater desalination. Resistance of the isolates to 10 antibiotics (Amikacin, Ampicillin, chloramphenicol, gentamycine, minocylin, nalidixicacid, neomycin, streptomycin, Tetracycline, Tobramycin, and Trimethoprim) was tested. Both fecal coliforms and enterococci were multiple resistant to 2-10 antibiotics. However, temperature variation during summer and winter did not affect resistance of the isolates to antibiotics. The significance of this investigation may be indicator to the environmental TSE pollution.

Keywords: antibiotic resistance, bacteria, environment, sewage treated effluent

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Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel A. Abugri, Robertson K. Boakai, Olufemi S. Ajayi

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The recalcitrance of pathogenic bacteria indicates that millions of people who are at risk of infection arising from chronic diseases, surgery, organ transplant, diabetes, and several other debilitating diseases present an aura of potentially untreatable illness due to resistance development. Antimicrobial resistance has successfully become a global health menace, and resistances are often acquired by bacteria through health-care-related incidence (HRI) orchestrated by multi-drug resistant (MDR) and extended drug-resistant pathogens (EDRP). To understand the mechanisms S. Typhimurium uses to resist CDB, we study the abundance of LPS modification, Ergosterols, Mysristic palmitic resistance, Oleic acid resistance of susceptible and resistant S. Typhimurium. Using qPCR, we also analyzed the expression of selected genes known for enabling resistance in S. Typhimurium. We found high abundance of LPS, Ergosterols, Mysristic palmitic resistance, Oleic acid resistance of and high expression of resistant genes in S. Typhimurium compared to the susceptible strain. LPS modification, Ergosterols, Mysristic palmitic resistance, Oleic acid and genes such as Fims, integrons, blaTEM are important indicators of resistance development of S. typhimurium.

Keywords: antimicrobials, resistance, Cannabidiol, Salmonella, blaTEM, fimA, Lipopolysaccharide, Ergosterols

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Authors: Beata Lachtara, Renata Szewczyk, Katarzyna Bielinska, Kinga Wieczorek, Jacek Osek

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Listeria monocytogenes is an important human and animal pathogen that causes foodborne outbreaks. The bacteria may be present in different types of food: cheese, raw vegetables, sliced meat products and vacuum-packed sausages, poultry, meat, fish. The most common method, which has been used for the investigation of genetic diversity of L. monocytogenes, is PFGE. This technique is reliable and reproducible and established as gold standard for typing of L. monocytogenes. The aim of the study was characterization by molecular serotyping and PFGE analysis of L. monocytogenes strains isolated from fresh fish and fish products in Poland. A total of 301 samples, including fresh fish (n = 129) and fish products (n = 172) were, collected between January 2014 and March 2016. The bacteria were detected using the ISO 11290-1 standard method. Molecular serotyping was performed with PCR. The isolates were tested with the PFGE method according to the protocol developed by the European Union Reference Laboratory for L. monocytogenes with some modifications. Based on the PFGE profiles, two dendrograms were generated for strains digested separately with two restriction enzymes: AscI and ApaI. Analysis of the fingerprint profiles was performed using Bionumerics software version 6.6 (Applied Maths, Belgium). The 95% of similarity was applied to differentiate the PFGE pulsotypes. The study revealed that 57 of 301 (18.9%) samples were positive for L. monocytogenes. The bacteria were identified in 29 (50.9%) ready-to-eat fish products and in 28 (49.1%) fresh fish. It was found that 40 (70.2%) strains were of serotype 1/2a, 14 (24.6%) 1/2b, two (4.3%) 4b and one (1.8%) 1/2c. Serotypes 1/2a, 1/2b, and 4b were presented with the same frequency in both categories of food, whereas serotype 1/2c was detected only in fresh fish. The PFGE analysis with AscI demonstrated 43 different pulsotypes; among them 33 (76.7%) were represented by only one strain. The remaining 10 profiles contained more than one isolate. Among them 8 pulsotypes comprised of two L. monocytogenes isolates, one profile of three isolates and one restriction type of 5 strains. In case of ApaI typing, the PFGE analysis showed 27 different pulsotypes including 17 (63.0%) types represented by only one strain. Ten (37.0%) clusters contained more than one strain among which four profiles covered two strains; three had three isolates, one with five strains, one with eight strains and one with ten isolates. It was observed that the isolates assigned to the same PFGE type were usually of the same serotype (1/2a or 1/2b). The majority of the clusters had strains of both sources (fresh fish and fish products) isolated at different time. Most of the strains grouped in one cluster of the AscI restriction was assigned to the same groups in ApaI investigation. In conclusion, PFGE used in the study showed a high genetic diversity among L. monocytogenes. The strains were grouped into varied clonal clusters, which may suggest different sources of contamination. The results demonstrated that 1/2a serotype was the most common among isolates from fresh fish and fish products in Poland.

Keywords: Listeria monocytogenes, molecular characteristic, PFGE, serotyping

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Authors: Gajanan V. Mane, Rashmi More, Mahesh S. Sonawane, Tushar Lodha, Rohit Sharma

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The diversity of fungi isolated from two different termite species viz., Odontoterms assmuthi and O. abesus was investigated by dilution- plate method, combined with morphological characteristics and sequencing of internal transcribed spacer region. In total, ninety-six fungi were isolated and purified, out of which 69 isolates were obtained from O. assmuthi belonging to 18 genera and 31 species, whereas 27 isolates were obtained from O. abesus belonging to 15 genera and 17 species. The fungal strains were screened for laccase, amylase, cellulase and pectinase enzymes production. Twenty-seven strains were positive for laccase, 59 strains were positive for amylase, 71 strains were positive for cellulase and 72 strains were positive for pectinase enzymes. The antimicrobial activities of the isolated fungi were tested by the dual plate culture method against standard pathogens. Bioactive secondary metabolites were identified by HPLC and LCMS. Four isolates viz., Penicillium goetzii MG 57, Epicoccum sp. MG 39, Penicillium tanzanicum MG 30, Aspergillus polyporicola MG 54, showed positive antimicrobial activity against standard pathogens, Streptococcus pneumonia MCC 2425, Staphylococcus aureus MCC 2408, Pseudomonas aeruginosa MCC 2080, Escherichia coli MCC 2412, Enterococcus faecalis MCC 2409, Klebsiella pneumonia MCC 2451, Micrococcus luteus MCC 2155 and Candida albicans MCC 1151. In conclusion, the study showed that the insect gut harbor fungal diversity, which is futuristic with biotechnological potential and could be a good source of enzymes and antibiotics.

Keywords: termites, fungi, its, enzyme, antimicrobial activity

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Authors: Farideh Tabatabaee Yazdi, Fereshteh Falah, Alireza Vasiee

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Introduction: Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is widely present in organisms. GABA is a kind of pharmacological and biological component and its application is wide and useful. Several important physiological functions of GABA have been characterized, such as neurotransmission and induction of hypotension. GABA is also a strong secretagogue of insulin from the pancreas and effectively inhibits small airway-derived lung adenocarcinoma and tranquilizer. Many microorganisms can produce GABA, and lactic acid bacteria have been a focus of research in recent years because lactic acid bacteria possess special physiological activities and are generally regarded as safe. Among them, the Lb. Brevis produced the highest amount of GABA. The major factors affecting GABA production have been characterized, including carbon sources and glutamate concentration. The use of food industry waste to produce valuable products such as amino acids seems to be a good way to reduce production costs and prevent the waste of food resources. In a dairy factory, a high volume of sludge is produced from a separator that contains useful compounds such as growth factors, carbon, nitrogen, and organic matter that can be used by different microorganisms such as Lb.brevis as carbon and nitrogen sources. Therefore, it is a good source of GABA production. GABA is primarily formed by the irreversible α-decarboxylation reaction of L-glutamic acid or its salts, catalysed by the GAD enzyme. In the present study, this aim was achieved for the fast-growing of Lb.brevis and producing GABA, using the dairy industry sludge as a suitable growth medium. Lactobacillus Brevis strains obtained from Microbial Type Culture Collection (MTCC) were used as model strains. In order to prepare dairy sludge as a medium, sterilization should be done at 121 ° C for 15 minutes. Lb. Brevis was inoculated to the sludge media at pH=6 and incubated for 120 hours at 30 ° C. After fermentation, the supernatant solution is centrifuged and then, the GABA produced was analyzed by the Thin Layer chromatography (TLC) method qualitatively and by the high-performance liquid chromatography (HPLC) method quantitatively. By increasing the percentage of dairy sludge in the culture medium, the amount of GABA increased. Also, evaluated the growth of bacteria in this medium showed the positive effect of dairy sludge on the growth of Lb.brevis, which resulted in the production of more GABA. GABA-producing LAB offers the opportunity of developing naturally fermented health-oriented products. Although some GABA-producing LAB has been isolated to find strains suitable for different fermentations, further screening of various GABA-producing strains from LAB, especially high-yielding strains, is necessary. The production of lactic acid, bacterial gamma-aminobutyric acid, is safe and eco-friendly. The use of dairy industry waste causes enhanced environmental safety. Also provides the possibility of producing valuable compounds such as GABA. In general, dairy sludge is a suitable medium for the growth of Lactic Acid Bacteria and produce this amino acid that can reduce the final cost of it by providing carbon and nitrogen source.

Keywords: GABA, Lactobacillus, HPLC, dairy sludge

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Authors: Mohammadreza Hajialigol, Nargues Falahi Charkhabi, Fatemeh Shahryari, Saadat Sarikhani

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BACKGROUND AND OBJECTIVES Walnut canker is characterized by brown to blackish roundish blotches on the trunks and main branches, necrosis of inner bark and bleeding with dark brown to black-colored exudates. The present study aimed to identify the causative agents of walnut decline by their phenotypic features, approval of pathogenicity, the partial sequencing of the housekeeping genes in Razavi Khorasan. MATERIAL AND METHODS Ten Symptomatic samples were collected from walnut orchards of Razavi Khorasan in 2019. Pathogenicity of all isolated strains was carried out on walnut immature fruits cv. ‘Hartley’ and young green twigs of cv. ‘Chandler’. All pathogenic strains were subjected to physiological, morphological and biochemical tests. 16S rRNA and housekeeping genes (fusA, leuS, and pyrG) were partially amplified and sequenced. RESULTS Eight strains were able to cause necrosis and a dark-colored region in the mesocarp of immature walnut fruits, and three representative strains caused necrosis on young inoculated twigs. Strains utilized starch, however, did not utilized esculin, Tween 20, Tween 80, and gelatin. The partial 16S rRNA gene sequence of strain KH7 indicated 99.63 % similarity to that of Citrobacter braakii ATCC5113T. The phylogenetic analyses based on the partial sequencing of three housekeeping genes, fusA (633 bp), pyrG (305), and leuS (640 bp), demonstrated that strains KH1, KH3, and KH7 belong to C. braakii species in a monophyletic clade with high bootstrap support. CONCLUSION To the best of our knowledge, this is the first report of C. braakii as a new plant pathogen which cause walnut decline. Identification of bacteria associated with walnut decline will eventually improve our understanding of the etiology of the disease and may result in improved management techniques for control.

Keywords: emerging pathogens, Iran, juglans regia, MLSA

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Authors: Yu Pu, Chien-Ping Hsiao, Chien-Chang Huang, Chieh-Lun Cheng

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Free radicals can accelerate aging on human skin by causing lipid oxidation, protein denaturation, and even DNA mutation. Substances with the ability of anti-free radicals can be used as functional components in cosmetic products. Research are attracted to develop new anti-free radical components for cosmetic application. This study was aimed to evaluate the microbial metabolites on free radical scavenging ability. Two microorganisms, PU-01 and PU-02, were isolated from soil of hot spring environment and grew in LB agar at 50°C for 24 h. The suspension was collected by centrifugation at 4800 g for 3 min, The anti-free radical activity was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging assay. The result showed that the growth medium of PU-01 presented a higher DPPH scavenging effect than that of PU-02. This study presented potential anti-free radical components from microbial metabolites that might be applied in anti-aging cosmetics.

Keywords: anti-ageing, anti-free radical, biotechnology, microorganism

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Authors: Abu Salim Mustafa

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Brucellosis is a zoonotic disease endemic in Kuwait. Human brucellosis can be caused by several Brucella species with Brucella melitensis causing the most severe and Brucella abortus the least severe disease. Furthermore, relapses are common after successful chemotherapy of patients. The classical biochemical methods of culture and serology for identification of Brucellae provide information about the species and serotypes only. However, to differentiate between relapse and reinfection/epidemiological investigations, the identification of genotypes using molecular methods is essential. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-16] were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. The 16S rRNA gene sequencing suggested that all the strains were B. melitensis and real-time PCR confirmed their species identity as B. melitensis. The ERIC-PCR band profiles produced a dendrogram of 75 branches suggesting each strain to be of a unique type. The cluster classification, based on ~ 80% similarity, divided all the ERIC genotypes into two clusters, A and B. Cluster A consisted of 9 ERIC genotypes (A1-A9) corresponding to 9 individual strains. Cluster B comprised of 13 ERIC genotypes (B1-B13) with B5 forming the largest cluster of 51 strains. MLVA-16 identified all isolates as B. melitensis and divided them into 71 MLVA-types. The cluster analysis of MLVA-16-types suggested that most of the strains in Kuwait originated from the East Mediterranean Region, a few from the African group and one new genotype closely matched with the West Mediterranean region. In conclusion, this work demonstrates that B. melitensis, the most pathogenic species of Brucella, is prevalent in Kuwait. Furthermore, MLVA-16 is the best molecular method, which can identify the Brucella species and genotypes as well as determine their origin in the global context. Supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: Brucella, ERIC-PCR, MLVA-16, RT-PCR, 16S rRNA gene sequencing

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Authors: Wisnu Adi Yulianto, Chatarina Lilis Suryani, Mamilisti Susiati, Hendy Indra Permana

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Parboiled rice was developed to produce rice that has low glycemic index, especially for diabetics. Yet, parboiled rice is not enough because diabetics also lack of chromium. The sign of chromium (Cr) deficiency in diabetics is impaired glucose tolerance. Cr fortification was done for increasing Cr content in rice. Naturally-occurring compounds that have been proven to improve insulin sensitivity include Cr and polyphenol found in cinnamon, pandan and bay leaf. This research aimed to evaluate content of resistant starch and glycemic index of Cr - fortified - parboiled rice (Cr-PR) coated with herbal extracts. Variety of unhulled rice and forticant used in the experiment were Ciherang and CrCl3, respectively. Three herbal extracts used were cinnamon, pandan and bay leaf. Each concentration of herbal extracts in the amount of 3%, 6%, and 9% were added in the coating substance to coat Cr-PR. Resistant starch (RS) content was determined by enzymatic process through glucooxydase method. Testing of the GI was conducted on 18 non-diabetic volunteers. RS content of Cr-PR coated with herbal extracts ranged between 8.27 – 8.84 % (dry weight). Cr-PR coated with all herbal extracts of 3% concentration had higher RS content than the ones with herbal extracts of 6% and 9% concentration (P <0.05). Value of the rice GI ranged 29 - 40. The lowest GI (29-30) was attained by the rice coated with enrichment of 6-9% cinnamon extract.

Keywords: coating, Cr-fortified-parboiled rice, glycemic index, herbal extracts, resistant starch

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Authors: Tomokazu Shirai, Akihiko Kondo

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Technologies of metabolic engineering or synthetic biology are essential for effective microbial bioproduction. It is especially important to develop an in silico tool for designing a metabolic pathway producing an unnatural and valuable chemical such as fossil materials of fuel or plastics. We here demonstrated two in silico tools for designing novel metabolic pathways: BioProV and HyMeP. Furthermore, we succeeded in creating an artificial metabolic pathway by enzyme engineering.

Keywords: bioinformatics, metabolic engineering, synthetic biology, genome scale model

Procedia PDF Downloads 316

Authors: Fnu Asina, Ivana Brzonova, Keith Voeller, Yun Ji, Alena Kubatova, Evguenii Kozliak

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Lignin, a heterogeneous three-dimensional biopolymer, is one of the building blocks of lignocellulosic biomass. Due to its limited chemical reactivity, lignin is currently processed as a low-value by-product in pulp and paper mills. Among various industrial lignins, Kraft lignin represents a major source of by-products generated during the widely employed pulping process across the pulp and paper industry. Therefore, valorization of Kraft lignin holds great potential as this would provide a readily available source of aromatic compounds for various industrial applications. Microbial degradation is well known for using both highly specific ligninolytic enzymes secreted by microorganisms and mild operating conditions compared with conventional chemical approaches. In this study, the degradation of Indulin AT lignin was assessed by comparing the effects of Basidiomycetous fungi (Coriolus versicolour and Trametes gallica) and Actinobacteria (Mycobacterium sp. and Streptomyces sp.) to two commercial laccases, T. versicolour ( ≥ 10 U/mg) and C. versicolour ( ≥ 0.3 U/mg). After 54 days of cultivation, the extent of microbial degradation was significantly higher than that of commercial laccases, reaching a maximum of 38 wt% degradation for C. versicolour treated samples. Lignin degradation was further confirmed by thermal carbon analysis with a five-step temperature protocol. Compared with commercial laccases, a significant decrease in char formation at 850ºC was observed among all microbial-degraded lignins with a corresponding carbon percentage increase from 200ºC to 500ºC. To complement the carbon analysis result, chemical characterization of the degraded products at different stages of the delignification by microorganisms and commercial laccases was performed by Pyrolysis-GC-MS.

Keywords: lignin, microbial degradation, pyrolysis-GC-MS, thermal carbon analysis

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

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

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

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