Search results for: fungal pathogens
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 813

Search results for: fungal pathogens

273 Sensitivity of Acanthamoeba castellanii-Grown Francisella to Three Different Disinfectants

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

Abstract:

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

Procedia PDF Downloads 73
272 Inactivation and Stress Response of Salmonella enterica Serotype Typhimurium lt21 upon Cold Gas-Phase Plasma Treatment

Authors: Zoran Herceg, Tomislava Vukušić, Anet Režek Jambrak, Višnja Stulić

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Today one of the greatest challenges are directed to the safety of food supply. If food pathogens are ingested they can cause human illnesses. Because of that new technologies that are effective in microbial reduction are developing to be used in food industries. One of such technology is cold gas phase plasma. Salmonella enterica was studied as one of the pathogenes that can be found in food. The aim of this work was to examine the inactivation rate and stress response of plasma treated cells of Salmonella enterica inoculated in apple juice. After the treatment cellular leakage, phenotypic changes in plasma treated cells-biofilm formation and degree of recovery were conducted. Sample volume was inoculated with 5 mL of pure culture of Salmonella enterica and 15 mL of apple juice. Statgraphics Centurion software (StatPoint Technologies, Inc., VA, USA) was used for experimental design and statistical analyses. Treatment time (1, 3, 5 min) and gas flow (40, 60, 80 L/min) were changed. Complete inactivation and 0 % of recovery after the 48 h was observed at these experimental treatments: 3 min; 40 L/min, 3 min; 80 L/min, 5 min; 40 L/min. Biofilm reduction was observed at all treated samples. Also, there was an increase in cellular leakage with a longer plasma treatment. Although there were a significant reduction and 0 % of recovery after the plasma treatments further investigation of the method is needed to clarify whether there are sensorial, physical and chemical changes in juices after the plasma treatment. Acknowledgments: The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for the preservation of liquid foods'.

Keywords: salmonella enterica serotype typhimurium lt21, gas-phase plasma treatment, inactivation, stress response

Procedia PDF Downloads 283
271 Isolation and Biological Activity of Betulinic and Oleanolic Acids from the Aerial Plant Parts of Maesobotrya Barteri (Baill)

Authors: Christiana Ene Ogwuche, Joseph Amupitan, George Ndukwe, Rachael Ayo

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Maesobotrya barteri (Baill), belonging to the family Euphorbiaceae, is a medicinal plant growing widely in tropical Africa. The Aerial plant parts of Maesobotrya barteri (Baill) were collected fresh from Orokam, Ogbadibo local Government of Benue State, Nigeria in July 2013. Taxonomical identification was done by Mallam Musa Abdullahi at the Herbarium unit of Biological Sciences Department, ABU, Zaria, Nigeria. Pulverized aerial parts of Maesobotrya barteri (960g) was exhaustively extracted successively using petroleum ether, chloroform, ethyl acetate and methanol and concentrated in the rotary evaporator at 40°C. The Petroleum ether extract had the second highest activity against test microbes from preliminary crude microbial screenings. The Petroleum ether extract was subjected to phytochemical studies, antimicrobial analysis and column chromatography (CC). The column chromatography yielded fraction PE, which was further purified using preparative thin layer chromatography to give PE1. The structure of the isolated compound was established using 1-D NMR and 2-D NMR spectroscopic analysis and by direct comparison with data reported in literature was confirmed to be a mixture, an isomer of Betulinic acid and Oleanolic acid, both with the molecular weight (C₃₀H₄₈O₃). The bioactivity of this compound was carried out using some clinical pathogens and the activity compared with standard drugs, and this was found to be comparable with the standard drug.

Keywords: Maesobotrya barteri, medicinal plant, bioactivity, petroleum spirit extract, butellinic acid, oleanilic acid

Procedia PDF Downloads 167
270 Assessing the Seed Yield of Some Varieties of Sesame (Sesami indicum) Under Disease Condition (Cercospora Leaf Spot) Caused by (Cercospora sesami, Zimm) and Identifying Disease Resistant Varieties

Authors: P. S. Akami, H. Nahunnaro, A. Zubainatu

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Cercospora leaf spot (Cercospora sesami. Zimm) has been identified as one of the most prevalent diseases, posing serious constraints to sesame production in producing areas. Two sets of experiments were carried out. The first and second experiments were conducted in the Modibbo Adama University of Technology Yola at the Crop Production and Horticulture and Plant Science Departments, respectively. The field experiment was carried out using a Randomized Complete Block Design and was replicated three times on a plot size of 4m x 5m with four sesame varieties and three Mancob-M fungicide levels (0g, 2g and 4g) to give a total of Twelve treatments. The laboratory experiment involved the isolation of the pathogens from diseased leaves with symptoms of Cercospora leaf spot, which was identified as Cercospora sesami. Data collected were subjected to analysis of variance for a randomized complete block design using SAS (1999) statistical package. The treatment means that are significantly different were separated using the Least Significant Difference at P=0.05. The result revealed that 4g Mancob M recorded the lowest mean value for disease incidence and severity at 8WAS, which was 90.30% and 35.60%, respectively, while the control (0g) recorded the highest mean value for disease incidence and severity at 90.30% and 59.80% respectively. Ex-Sudan recorded the lowest value of 720 kg/ha, while NCRIBEN 03 recorded the highest yield of 834 kg/ha-¹. For the concentrations, 2g recorded a higher yield of 843 kg/ha-¹ followed by 0g, which recorded 765 kg/ha-¹. Conclusively, Cercospora leaf spot of sesame was found to be prevalent. E8 has a higher resistance to the disease, while NCRIBEN 03 tends to be more susceptible. It is therefore recommended that further trials should be carried out using different varieties in different locations.

Keywords: disease, evaluation, prevalence, treatment, resistance

Procedia PDF Downloads 49
269 Low Temperature Biological Treatment of Chemical Oxygen Demand for Agricultural Water Reuse Application Using Robust Biocatalysts

Authors: Vedansh Gupta, Allyson Lutz, Ameen Razavi, Fatemeh Shirazi

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The agriculture industry is especially vulnerable to forecasted water shortages. In the fresh and fresh-cut produce sector, conventional flume-based washing with recirculation exhibits high water demand. This leads to a large water footprint and possible cross-contamination of pathogens. These can be alleviated through advanced water reuse processes, such as membrane technologies including reverse osmosis (RO). Water reuse technologies effectively remove dissolved constituents but can easily foul without pre-treatment. Biological treatment is effective for the removal of organic compounds responsible for fouling, but not at the low temperatures encountered at most produce processing facilities. This study showed that the Microvi MicroNiche Engineering (MNE) technology effectively removes organic compounds (> 80%) at low temperatures (6-8 °C) from wash water. The MNE technology uses synthetic microorganism-material composites with negligible solids production, making it advantageously situated as an effective bio-pretreatment for RO. A preliminary technoeconomic analysis showed 60-80% savings in operation and maintenance costs (OPEX) when using the Microvi MNE technology for organics removal. This study and the accompanying economic analysis indicated that the proposed technology process will substantially reduce the cost barrier for adopting water reuse practices, thereby contributing to increased food safety and furthering sustainable water reuse processes across the agricultural industry.

Keywords: biological pre-treatment, innovative technology, vegetable processing, water reuse, agriculture, reverse osmosis, MNE biocatalysts

Procedia PDF Downloads 101
268 Investigating the Combined Medicinal Effects of Withania Somnifera (Ashwaghandha) and Murraya Koenigii (Curry Pata) in Vitro

Authors: Sadia Roshan, Kulsoom Sughra, Shazia Shamas, Shamaila Irum, Haleema Sadia

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To evaluate synergistic medicinal effects of Withania somnifera (Ashwaghandha) and Murraya koenigii (Curry pata) in vitro. Antimicrobial activity was determined using the disc diffusion method against five bacterial and two fungal strains. The antioxidant activity was evaluated by the DPPH assay. The antidiabetic activity was evaluated by alpha-glucosidase inhibition assay and alpha-amylase inhibition assay. Synergistic antibacterial activity was observed against all the strains of bacteria, either Gram-positive or Gram-negative and fungi under study conditions. The maximum antibacterial activity was displayed by combined extract against E. coli i.e. 26±0.4mm. Maximum antifungal activity was shown by combined extract against Aspergillus niger, i.e., 17.3±0.5mm. The antioxidant activity of the combined extract was also significant. Alpha-glucosidase inhibition and alpha-amylase inhibition assays also showed synergism. Results indicate that Withania somnifera and Murraya koengii have medicinal properties. The combined extract of both plants is more potent than their individual extracts, suggesting that these can work in synergism. The research suggests that different plant extracts could be used in combination to increase their medicinal activities by many folds, thus giving an insight into future use of herbal medication.

Keywords: withania somnifera, murraya koenigii, antimicrobial activity, gram-positive bacetria, gram-negative bacteria

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267 Susceptibility Assessment and Genetic Diversity of Iranian and CIMMYT Wheat Genotypes to Common Root Rot Disease Bipolaris sorokiniana

Authors: Mehdi Nasr Esfahani, Abdal-Rasool Gholamalian, Abdelfattah A. Dababat

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Wheat, Triticum aestivum L. is one of the most important and strategic crops in the human diet. Several diseases threaten this particular crop. Common root rot disease of wheat by a fungal agent, Bipolaris sorokiniana is one of the important diseases, causing considerable losses worldwide. Resistant sources are the only feasible and effective method of control for managing diseases. In this study, the response of 33 domestic and exotic wheat genotypes, including cultivars and promising lines were screened to B. sorokiniana at greenhouse and field conditions, based on five scoring scale indexes of 0 to 100 severity percentage. The screening was continued on resistant wheat genotypes and repeated several times to confirm the greenhouse and field results. Statistical and cluster analysis of data was performed using SAS and SPSS software, respectively. The results showed that, the response of wheat genotypes to the disease in the greenhouse and field conditions was highly significant. The highest rate of common root rot disease infection, B. sorokiniana in the greenhouse and field, was of CVS. Karkheh and Beck Cross-Roshan with 60.83% and 59.16% disease severity respectively, and the lowest one were in cv. Alvand with 18.33%, followed by cv. Baharan with 19.16% disease severity, with a highly significant difference respectively. The remaining wheat genotypes were located in between these two highest and lowest infected groups to B. sorokiniana significantly. There was a high correlation coefficient between the related statistical groups and cluster analysis.

Keywords: wheat, rot, root, crown, fungus, genotype, resistance

Procedia PDF Downloads 108
266 Multiple Etiologies and Incidences of Co-Infections in Childhood Diarrhea in a Hospital Based Screening Study in Odisha, India

Authors: Arpit K. Shrivastava, Nirmal K. Mohakud, Subrat Kumar, Priyadarshi S. Sahu

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Acute diarrhea is one of the major causes of morbidity and mortality among children less than five years of age. Multiple etiologies have been implicated for infectious gastroenteritis causing acute diarrhea. In our study fecal samples (n=165) were collected from children (<5 years) presenting with symptoms of acute diarrhea. Samples were screened for viral, bacterial, and parasitic etiologies such as Rotavirus, Adenovirus, Diarrhoeagenic Escherichia coli (EPEC, EHEC, STEC, O157, O111), Shigella spp., Salmonella spp., Vibrio cholera, Cryptosporidium spp., and Giardia spp. The overall results from our study showed that 57% of children below 5 years of age with acute diarrhea were positive for at least one infectious etiology. Diarrhoeagenic Escherichia coli was detected to be the major etiological agent (29.09%) followed by Rotavirus (24.24%), Shigella (21.21%), Adenovirus (5.45%), Cryptosporidium (2.42%), and Giardia (0.60%). Among the different DEC strains, EPEC was detected significantly higher in <2 years children in comparison to >2 years age group (p =0.001). Concurrent infections with two or more pathogens were observed in 47 of 160 (28.48%) cases with a predominant incidence particularly in <2-year-old children (66.66%) compared to children of 2 to 5 years age group. Co-infection of Rotavirus with Shigella was the most frequent combination, which was detected in 17.94% cases, followed by Rotavirus with EPEC (15.38%) and Shigella with STEC (12.82%). Detection of multiple infectious etiologies and diagnosis of the right causative agent(s) can immensely help in better management of acute childhood diarrhea. In future more studies focusing on the detection of cases with concurrent infections must be carried out, as we believe that the etiological agents might be complementing each other’s strategies of pathogenesis resulting in severe diarrhea.

Keywords: children, co-infection, infectious diarrhea, Odisha

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265 Cytotoxic Activity of Marine-derived Fungi Trichoderma Longibrachiatum Against PANC-1 Cell Lines

Authors: Elin Julianti, Marlia Singgih, Masayoshi Arai, Jianyu Lin, Masteria Yunovilsa Putra, Muhammad Azhari, Agnia S. Muharam

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The search for a source of new medicinal compounds with anticancer activity from natural products has become important to resolve the ineffectiveness problem of pancreatic cancer therapy. Fungal marine microorganisms are prolific sources of bioactive natural products. In this present study, the ethyl acetate extract of cultured broth of Trichoderma longibrachiatum marine sponge-derived fungi exhibited selective cytotoxicity against human pancreatic carcinoma PANC-1 cells cultured under glucose-deficient conditions (IC50 = 98,4 µg/mL). The T. longibrachiatum was fermented by the static method at room temperature for 60 days. The culture broth was extracted using ethyl acetate by liquid-liquid extraction method. The liquid-liquid extraction was conducted toward the ethyl extract by using 90% MeOH-H₂O and n-|Hexane as a solvent. The extract of 90% MeOH-H₂O was fractionated by liquid extraction using by C₁₈ reversed-phase vacuum flash chromatography using mixtures of MeOH-H₂O, from 50:50 to 100:0, and 1% TFA MeOH as the eluents to yield six fractions. The fraction 2 (MeOH-H2O, 70:30) and fraction 3 (MeOH-H2O, 80:20) showed moderate cytotoxicity with IC50 value of 119.3 and 274.7 µg/mL, respectively. Fraction 4 (MeOH-H₂O, 90:10) showed the highest cytotoxicity activity with IC₅₀value of < 10 µg/mL. The chemical compounds of the fractions that are responsible for cytotoxic activity are potent for further investigation.

Keywords: cytotoxic activity, trichoderma longibrachiatum, marine-derived fungi, PANC-1 cell line

Procedia PDF Downloads 259
264 Isolation and Identification Fibrinolytic Protease Endophytic Fungi from Hibiscus Leaves in Shah Alam

Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

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Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

Procedia PDF Downloads 383
263 M. J. Rodríguez, F. M. Sánchez, B. Velardo, P. Calvo, M. J. Serradilla, J. Delgado, J. M. López

Authors: Q. Rzina, M. Lahrouni, S. Rida, N. Saadaoui, Y. Almossaid, K. Oufdou, K. Fares

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Many organic solid wastes are produced in the world. Poultry manure (PM), municipal organic wastes (MOW) and sugar beet lime sludge (LS) are produced in large quantities in Morocco. The co-composting of these organic wastes was investigated. The recycling and the valorization of such wastes is environmentally and economically beneficial especially for PM which is known source of bacterial pathogens. The aerobic biodegradation process was carried out by using three windrows of variable compositions: C1 prepared without LS (only MOW were composted with PM), C2 prepared from MOW plus PM and10% LS; and the last one C3 from MOW plus PM and 20% LS. The main process physico-chemical parameters (temperature, pH, humidity and C/N) and microbiological populations (mesophilic and thermophilic flora, total coliform, fecal coliform, Streptococci, Staphylococcus aureus and mesophilic fungi) were monitored over three months to ascertain the compost maturity and to ensure the compost hygienic aspect. The final products were characterized by their relatively high organic matter content, and low C/N ratio of 10.6-10.9. The organic matter degradation was reached approximately 59% for C2 and C3. In addition, the monitoring of the microbial population showed that the produced composts are mature and hygienic. The agronomic valorization of the final composts was tested on radish plant with tree level of composts and poultry manure without composting. The primary results of field trial showed a growth of radish plant biomass and root development without any phytotoxicity detected which reflects the quality of the composts produced. As for poultry manure it allowed to have a better results than other composts because of its readily available nitrogen.

Keywords: compost, municipal organic wastes, poultry manure, radish crop, sugar beet lime sludge

Procedia PDF Downloads 279
262 Difference in Virulence Factor Genes Between Transient and Persistent Streptococcus Uberis Intramammary Infection in Dairy Cattle

Authors: Anyaphat Srithanasuwan, Noppason Pangprasit, Montira Intanon, Phongsakorn Chuammitri, Witaya Suriyasathaporn, Ynte H. Schukken

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Streptococcus uberis is one of the most common mastitis-causing pathogens, with a wide range of intramammary infection (IMI) durations and pathogenicity. This study aimed to compare shared or unique virulence factor gene clusters distinguishing persistent and transient strains of S. uberis. A total of 139 S. uberis strains were isolated from three small-holder dairy herds with a high prevalence of S. uberis mastitis. The duration of IMI was used to categorize bacteria into two groups: transient and persistent strains with an IMI duration of less than 1 month and longer than 2 months, respectively. Six representative S. uberis strains, three from each group (transience and persistence) were selected for analysis. All transient strains exhibited multi-locus sequence types (MLST), indicating a highly diverse population of transient S. uberis. In contrast, MLST of persistent strains was available in an online database (pubMLST). Identification of virulence genes was performed using whole-genome sequencing (WGS) data. Differences in genomic size and number of virulent genes were found. For example, the BCA gene or alpha-c protein and the gene associated with capsule formation (hasAB), found in persistent strains, are important for attachment and invasion, as well as the evasion of the antimicrobial mechanisms and survival persistence, respectively. These findings suggest a genetic-level difference between the two strain types. Consequently, a comprehensive study of 139 S. uberis isolates will be conducted to perform an in-depth genetic assessment through WGS analysis on an Illumina platform.

Keywords: Streptococcus Uberis, mastitis, whole genome sequence, intramammary infection, persistent S. Uberis, transient s. Uberis

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261 Survey of Potato Viral Infection Using Das-Elisa Method in Georgia

Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

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Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

Procedia PDF Downloads 251
260 The Role of a Novel DEAD-Box Containing Protein in NLRP3 Inflammasome Activation

Authors: Yi-Hui Lai, Chih-Hsiang Yang, Li-Chung Hsu

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The inflammasome is a protein complex that modulates caspase-1 activity, resulting in proteolytic cleavage of proinflammatory cytokines such as IL-1β and IL-18, into their bioactive forms. It has been shown that the inflammasomes play a crucial role in the clearance of pathogenic infection and tissue repair. However, dysregulated inflammasome activation contributes to a wide range of human diseases such as cancers and auto-inflammatory diseases. Yet, regulation of NLRP3 inflammasome activation remains largely unknown. We discovered a novel DEAD box protein, whose biological function has not been reported, not only negatively regulates NLRP3 inflammasome activation by interfering NLRP3 inflammasome assembly and cellular localization but also mitigate pyroptosis upon pathogen evasion. The DEAD-box protein is the first DEAD-box protein gets involved in modulation of the inflammasome activation. In our study, we found that caspase-1 activation and mature IL-1β production were largely enhanced upon LPS challenge in the DEAD box-containing protein- deleted THP-1 macrophages and bone marrow-derived macrophages (BMDMs). In addition, this DEAD box-containing protein migrates from the nucleus to the cytoplasm upon LPS stimulation, which is required for its inhibitory role in NLRP3 inflammasome activation. The DEAD box-containing protein specifically interacted with the LRR motif of NLRP3 via its DEAD domain. Furthermore, due to the crucial role of the NLRP3 LRR domain in the recruitment of NLRP3 to mitochondria and binding to its adaptor ASC, we found that the interaction of NLRP3 and ASC was downregulated in the presence of the DEAD box-containing protein. In addition to the mechanical study, we also found that this DEAD box protein protects host cells from inflammasome-triggered cell death in response to broad-ranging pathogens such as Candida albicans, Streptococcus pneumoniae, etc., involved in nosocomial infections and severe fever shock. Collectively, our results suggest that this novel DEAD box molecule might be a key therapeutic strategy for various infectious diseases.

Keywords: inflammasome, inflammation, innate immunity, pyroptosis

Procedia PDF Downloads 256
259 Contribution to the Production of Phenazine Antibiotics Effect Type Compounds by Some Strains of Pseudomonas spp.fluorescent

Authors: Nacéra Benoussaid, Lehalali Meriem, Benchabane Messaoud

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Our work focuses on the production of compound antibiotic effect of volatile nature namely hydrogen cyanide and the production and identification of molecules phénazinique by some strains of fluorescent Pseudomonas spp isolated from the rhizosphere of some trees for a possible use as bio pesticides antifungal effect and/or antibiotic. We tested the production of hydrogen cyanide of 21 strains of Pseudomonas spp. fluorescent among them 19 strains (90, 47%) showed a positive cyanogenesis.The antagonism test executed in vitro showed that Pseudomonas strains have a higher anti fungal effect relative to their antibacterial effect with diameters of inhibition zones up to 3, 9 cm recorded by the strain F48 against Coleosporiumsp compared with recorded results against bacteria with a maximum inhibition of 1, 26 cm among this antagonistic strain.Three strains were selected by testing for producing phénazines namely PI9, BB9 and F20. The effect of the antimicrobial activity was performed on different culture media (GN, King B, ISP2 and PDA). The results of our study allowed us to retain the King B medium as ideal medium for the production of secondary metabolite. The produced phenazinique compounds was extracted from various organic solvents, and after the results of antibiographie against germs - targets, the extracts of ethyl acetate gave the best results compared to dichloromethane and hexane.The Analysis of these compounds of antibiotic phenazinique effect within layer chromatography (CCM) and high performance liquid chromatography( HPLC) indicate that both strains PI9 and F20 are productive of phenazine-1-carboxylic acid (PCA). The BB9 strain is suspected to be productive of another phenazinique compound.

Keywords: Pseudomonas ssp. fluorescents, antagonism in vitro, secondary metabolite, phenazines, biopesticide.

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258 Diverse Sensitivity to Ultraviolet Radiation of DNA and RNA Viruses

Authors: Nickolay Nosik, Dmitry Nosik, Marina Bochkova, Nina Kondrashina, Olga Lobach

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The bactericidal effect of UV radiation is known for long time and widely used for inactivation of pathogens but for viruses it is not so uniform. Due to a wide variety of viruses their sensitivity to UV radiation is quite different and not quite predictable. The goal of the study was to determine the inactivation kinetics of UV radiation ( 254 nm) of the viruses of social importance (HIV), as well as test-viruses (poliovirus, adenovirus) used for the evaluation of the viral inactivation efficacy of germicides. Methods: DNA viruses- adenovirus, type 5; Herpes simplex virus (HSV), type 1, and RNA viruses–human immunodeficiency virus (HIV), type 1 and poliovirus, type 1 (Sabin strain) were obtained from State collection of viruses ( The D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15-watt low-pressure mercury vapor lamp (over 60% 254nm). The samples of 5cm2 were placed direct under the UV lamp flow (h-0.3m). Log reduction value was used as a marker for the rate of virus inactivation. Results: The data obtained indicate that poliovirus (one of the viruses most resistant to chemical germicides) and HSV are rather sensitive to UV radiation ( D90 =250-311 J/m2). Adenovirus is much more resistant to UV radiation (750 J/m2 ). The kinetics of adenovirus inactivation : 0 min- 5.0 lg TCID50, 10 min - 5,0, 15 min -4,0, 30 min – 3.5, 60 min – 1,0, 75 min -0,5 lg TCID50, 90 min –virus not detectable. HIV is most resistant to UV radiation among the studied viruses. It takes more than 4 hrs to inactivate the virus on the surface. D90 = 2000 J/m2 Conclusion: The results of the study show that there is no direct dependence between sensitivity to UV light and the size of the virion or presence\absence of the envelope of the virus. Poliovirus and adenovirus are small viruses (20-30nm poliovirus and 70-90nm adenovirus) and both are non-enveloped viruses but adenovirus 3-fold more resistant to UV radiation than poliovirus. It can be expected that viruses with more complicate structure, like Herpes virus (200nm) or HIV (80-100 nm), would be more sensitive to UV light. However, the very high resistance of HIV to UV radiation needs further investigation. The diverse resistance of the different viruses to UV radiation should be taken into the account when UV light is used to inactivate infectious viruses in hospitals and other public environments.

Keywords: HIV, HSV, inhibition of viruses, UV radiation

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257 Molecular Evolutionary Relationships Between O-Antigens of Enteric Bacteria

Authors: Yuriy A. Knirel

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Enteric bacteria Escherichia coli is the predominant facultative anaerobe of the colonic flora, and some specific serotypes are associated with enteritis, hemorrhagic colitis, and hemolytic uremic syndrome. Shigella spp. are human pathogens that cause diarrhea and bacillary dysentery (shigellosis). They are in effect E. coli with a specific mode of pathogenicity. Strains of Salmonella enterica are responsible for a food-borne infection (salmonellosis), and specific serotypes cause typhoid fever and paratyphoid fever. All these bacteria are closely related in respect to structure and genetics of the lipopolysaccharide, including the O-polysaccharide part (O‑antigen). Being exposed to the bacterial cell surface, the O antigen is subject to intense selection by the host immune system and bacteriophages giving rise to diverse O‑antigen forms and providing the basis for typing of bacteria. The O-antigen forms of many bacteria are unique, but some are structurally and genetically related to others. The sequenced O-antigen gene clusters between conserved galF and gnd genes were analyzed taking into account the O-antigen structures established by us and others for all S. enterica and Shigella and most E. coli O-serogroups. Multiple genetic mechanisms of diversification of the O-antigen forms, such as lateral gene transfer and mutations, were elucidated and are summarized in the present paper. They include acquisition or inactivation of genes for sugar synthesis or transfer or recombination of O-antigen gene clusters or their parts. The data obtained contribute to our understanding of the origins of the O‑antigen diversity, shed light on molecular evolutionary relationships between the O-antigens of enteric bacteria, and open a way for studies of the role of gene polymorphism in pathogenicity.

Keywords: enteric bacteria, O-antigen gene cluster, polysaccharide biosynthesis, polysaccharide structure

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256 A Study on the Microbilogical Profile and Antibiotic Sensitivity Pattern of Bacterial Isolates Causing Urinary Tract Infection in Intensive Care Unit Patients in a Tertiary Care Hospital in Eastern India

Authors: Pampita Chakraborty, Sukumar Mukherjee

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The study was done to determine the microbiological profile and changing pattern of the pathogens causing UTI in the ICU patients. All the patients admitted to the ICU with urinary catheter insertion for more than 48hours were included in the study. Urine samples were collected in a sterile container with aseptic precaution using disposable syringe and was processed as per standards. Antimicrobial susceptibility test was done by Disc Diffusion method as per CLSI guidelines. A total of 100 urine samples were collected from ICU patients, out of which 30% showed significant bacterial growth and 7% showed growth of candida spp. Prevalence of UTI was more in female (73%) than male (27.%). Gram-negative bacilli 26(86.67%) were more common in our study followed by gram-positive cocci 4(13.33%). The most common uropathogens isolated were Escherichia coli 14 (46.67%), followed by Klebsiella spp 7(23.33%), Staphylococcus aureus 4(13.33%), Acinetobacter spp 3(10%), Enterococcus faecalis 1(3.33%) and Pseudomonas aeruginosa 1(3.33%). Most of the Gram-negative bacilli were sensitive to amikacin (80%) and nitrofurantoin (80%), where as all gram-positive organisms were sensitive to Vancomycin. A large number ESBL producers were also observed in this study. The study finding showed that E.coli is the predominant pathogen and has increasing resistance pattern to the commonly used antibiotics. The study proposes that the adherence to antibiotic policy is the key ingredients for successful outcome in ICU patients and also emphasizes that repeated evaluation of microbial characteristics and continuous surveillance of resistant bacteria is required for selection of appropriate antibiotic therapy.

Keywords: antimicrobial sensitivity, intensive care unit, nosocomial infection, urinary tract infection

Procedia PDF Downloads 245
255 Development of an Aerosol Protection Capsule for Patients with COVID-19

Authors: Isomar Lima da Silva, Aristeu Jonatas Leite de Oliveira, Roberto Maia Augusto

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Biological isolation capsules are equipment commonly used in the control and prevention of infectious diseases in the hospital environment. This type of equipment, combined with pre-established medical protocols, contributes significantly to the containment of highly transmissible pathogens such as COVID-19. Due to its hermetic isolation, it allows more excellent patient safety, protecting companions and the health team. In this context, this work presents the development, testing, and validation of a medical capsule to treat patients affected by COVID-19. To this end, requirements such as low cost and easy handling were considered to meet the demand of people infected with the virus in remote locations in the Amazon region and/or where there are no ICU beds and mechanical ventilators for orotracheal intubation. Conceived and developed in a partnership between SAMEL Planos de Saúde and Instituto Conecthus, the device entitled "Vanessa Capsule" was designed to be used together with the NIV protocol (non-invasive ventilation), has an automatic exhaust system and filters performing the CO2 exchange, in addition to having BiPaps ventilatory support equipment (mechanical fans) in the Cabin Kit. The results show that the degree of effectiveness in protecting against infection by aerosols, with the protection cabin, is satisfactory, implying the consideration of the Vanessa capsule as an auxiliary method to be evaluated by the health team. It should also be noted that the medical observation of the evaluated patients found that the treatment against the COVID-19 virus started earlier with non-invasive mechanical ventilation reduces the patient's suffering and contributes positively to their recovery, in association with isolation through the Vanessa capsule.

Keywords: COVID-19, mechanical ventilators, medical capsule, non-invasive ventilation

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254 Inoculation of Cyanobacteria Improves the Lignin Content of Thymus vulgaris L.

Authors: Nasim Rasuli, Akram Ahmadi, Hossein Riahi, Zeinab Shariatmadari, Majid Ghorbani Nohooji, Pooyan Mehraban Joubani

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Cyanobacteria are one of the most promising sources of new biostimulants and have received much attention due to their diverse applications in biotechnology. These microorganisms enhance the growth and productivity of plants by producing plant growth stimulants and fixing atmospheric nitrogen. Thymus vulgaris L., a valuable medicinal plant from the Lamiaceae family, is widely distributed across the globe. essential oil of T. vulgaris is best characterized by the prominence of phenols, making them the key compounds in its composition. Lignin biosynthesis as a natural plant polyphenol plays a crucial role in promoting plant growth, strengthening cell walls, and increasing resistance to pathogens. In this study, the bioelicitor activity of five cyanobacterial suspensions including Anabaena torulosa ISB213, Nostoc calcicola ISB215, Nostoc ellipsosporum ISB217, Trichormus doliolum ISB214, and Oscillatoria sp. ISB2116 on the lignin content of the T. vulgaris L. was investigated. Pot experiments were performed by inoculation of a %2 algal extract to the soil of treated plants one week before planting and then every 20 days. After four months, the lignin content in the leaves of both treated and control plants was evaluated. The results demonstrated that the application of cyanobacteria significantly increased the lignin content in the leaves of treated plants compared to the control. The treatment with Oscillatoria sp. ISB216 and N. ellipsosporum ISB217 resulted in the highest lignin content, with an increase of 93.33% and 86.67%, respectively. These findings highlight the potential of cyanobacteria as bioelicitors, offering a viable alternative for enhancing the production of secondary metabolites in T. vulgaris. Consequently, this could contribute to the economic value of this medicinal plant.

Keywords: cyanobacteria, bioelicitor, thymus vulgaris, lignin

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253 Role of Giardia lamblia Infection in the Pathogenesis of Gastritis in Patients with Dyspepsia

Authors: Aly Kassem, Eman A. Sabet, Hanaa A. El-Hady, Doha S. Mohamed, Abeer Sheneef, Mona Fattouh, Mamdouh M. Esmat

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Objective: Giardia lamblia parasite is the most common protozoal infection in human. Concomitant Helecobacter Pylori (H. pylori) and Giardia lamblia infection is common for their similar mode of transmission and strong correlation to socioeconomic levels. Only few reports had described gastric giardiasis. Our aim was to detect H. pylori and Giardia in gastric antral mucosal biopsies from patients with dyspepsia. The impact of both pathogens on clinical, endoscopic and histopathogical changes was studied. Methods: 48 patients with dyspepsia (group1) and 28 control patients (patients undergoing esophagogastroduodenoscopy EGD for reasons other than dyspepsia), (group 2) were studied. Endoscopic data were reported and gastric biopsy specimens were obtained for subsequent PCR assay for both organisms and for histopathological and electron microscopic examination. Results: Endoscopic antral gastritis and duodenal lesions were found in both groups, however, they were significantly more frequently in group 1 (p= 0.002 and P= 0.0005 respectively). Esophageal lesions, nodular antral gastritis, gastric ulcers and superficial corpal gastritis were found only in group 1. PCR detected H. pylori infection in 58% Vs 64 % for group 1 and group 2 respectively (P: NS). Giardia infection was present in 67 % Vs 42 % for group 1 and group 2 respectively (P=0.0003, Odd ratio=2.6). Co-infection with H. pylori and Giardia was present in 33% of group 1 Vs 36% for group 2 (P:NS). Abnormal histologic findings were found in both groups, however, intestinal metaplasia was found in group 1 only. Cellular abnormalities in the form of cytoplasmic vacuoles, mitochondrial destruction or nuclear abnormalities were found by Electron microscopic study in infected subjects of both groups. Conclusion: H. pylori is not the only gastric pathogen in our community, gastric giardiasis is another pathogen. Its contribution might be a factor in persistent dyspepsia after H. pylori eradication.

Keywords: dyspepsia, gastritis, Giardia lamblia, H. pylori

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252 Ecological impacts of Cage Farming: A Case Study of Lake Victoria, Kenya

Authors: Mercy Chepkirui, Reuben Omondi, Paul Orina, Albert Getabu, Lewis Sitoki, Jonathan Munguti

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Globally, the decline in capture fisheries as a result of the growing population and increasing awareness of the nutritional benefits of white meat has led to the development of aquaculture. This is anticipated to meet the increasing call for more food for the human population, which is likely to increase further by 2050. Statistics showed that more than 50% of the global future fish diet will come from aquaculture. Aquaculture began commercializing some decades ago; this is accredited to technological advancement from traditional to modern cultural systems, including cage farming. Cage farming technology has been rapidly growing since its inception in Lake Victoria, Kenya. Currently, over 6,000 cages have been set up in Kenyan waters, and this offers an excellent opportunity for recognition of Kenya’s government tactic to eliminate food insecurity and malnutrition, create employment and promote a Blue Economy. However, being an open farming enterprise is likely to emit large bulk of waste hence altering the ecosystem integrity of the lake. This is through increased chlorophyll-a pigments, alteration of the plankton community, macroinvertebrates, fish genetic pollution, transmission of fish diseases and pathogens. Cage farming further increases the nutrient loads leading to the production of harmful algal blooms, thus negatively affecting aquatic and human life. Despite the ecological transformation, cage farming provides a platform for the achievement of the Sustainable Development Goals of 2030, especially the achievement of food security and nutrition. Therefore, there is a need for Integrated Multitrophic Aquaculture as part of Blue Transformation for ecosystem monitoring.

Keywords: aquaculture, ecosystem, blue economy, food security

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251 Production of Mycelial Biomass, Exopolysaccharide, and Enzyme during Solid-State Fermentation of Plant Raw Materials by Medicinal Mushrooms

Authors: Tamar Khardziani, Violeta Berikashvili, Amrosi Chkuaseli, Vladimir Elisashvili

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The main objectives of this proposal are to develop low-cost, innovative, and competitive technologies for the production of mycelial biomass of medicinal mushrooms as a natural food supplement for poultry. To fulfill this task, industrial strains of Lentinus edodes, Ganoderma lucidum, and Pleurotus ostreatus were used in this study. The solid-state fermentation (SSF) of wheat grains, wheat bran, and soy flour was performed in flasks and bags. Among nine mushroom strains, P. ostreatus 2191 appeared to be the most productive in protein biomass accumulation in the SSF of wheat bran. All mushrooms produced exopolysaccharide with the highest yield of 5-8 mg/mL depending on fungal strain and growth substrate. Supplementation of medium with 1% glycerol and 2-4% peptone favored mushroom growth and protein accumulation. Among inorganic nitrogen sources, KNO₃ also provided high biomass and protein production. The SSF of all growth substrates was accompanied by the secretion of cellulase and xylanase activities. The highest CMCase activity (12-13 U/g) was revealed in the cultivation of P. ostreatus 2191 using wheat bran as a growth substrate and ammonium sulfate or yeast extract as a nitrogen source, whereas the highest xylanase activity was detected in the fermentation of soy flour supplemented with peptone. Acknowledgments: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (Grant number STEM-22-2077).

Keywords: mushrooms, plant raw materials, fermentation, biomass protein, cellulase

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250 Culturable Microbial Diversity of Agave Artisanal Fermentations from Central Mexico

Authors: Thalía Moreno-García Malo, Santiago Torres-Ríos, María G. González-Cruz, María M. Hernández-Arroyo, Sergio R. Trejo-Estrada

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Agave atrovirens is the main source of agave sap, the raw material for the production of pulque, an artisanal fermented beverage, traditional since prehispanic times in the highlands of central Mexico. Agave sap is rich in glucose, sucrose and fructooligosaccharides, and strongly differs from agave syrup from A. tequilana, which is mostly a high molecular weight fructan. Agave sap is converted into pulque by a highly diverse microbial community which includes bacteria, yeast and even filamentous fungi. The bacterial diversity has been recently studied. But the composition of consortia derived from directed enrichments differs sharply from the whole fermentative consortium. Using classical microbiology methods, and selective liquid and solid media formulations, either bacterial or fungal consortia were developed and analyzed. Bacterial consortia able to catabolize specific prebiotic saccharides were selected and preserved for future developments. Different media formulations, selective for bacterial genera such as Bifidobacterium, Lactobacillus, Pediococcus, Lactococcus and Enterococcus were also used. For yeast, specific media, osmotic pressure and unique carbon sources were used as selective agents. Results show that most groups are represented in the enrichment cultures; although very few are recoverable from the whole consortium in artisanal pulque. Diversity and abundance vary among consortia. Potential bacterial probiotics obtained from agave sap and agave juices show tolerance to hydrochloric acid, as well as strong antimicrobial activity.

Keywords: Agave, pulque, microbial consortia, prebiotic activity

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249 Probiotic Antibacterial Test of Pediococcus pentosaceus Isolated from Dadih in Inhibiting Periodontitis Bacteria: In Vitro Study on Bacteria Aggregatibacter actinomycetemcomitans

Authors: Nurlaili Syafar Wulan, Almurdi, Suprianto Kosno

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Introduction: Periodontitis defined as an inflammatory disease of teeth supporting tissue with irritation of specific pathogens as the main aetiology. Periodontitis can be cured by giving medical action accompanied by administration of an antibiotic, but the use of antibiotic has a side effect that can cause bacterial resistance. This side effect can be corrected by probiotic, which has antibiotic-like substance but do not have bacterial resistance effect; it makes probiotic became a promising future periodontitis medication. West Sumatran people has their own typical traditional food product made from fermented buffalo’s milk called dadih, and it contained probiotics. Objectives: The aim of this study was to determine the ability of probiotic Pediococcus pentosaceus isolated from dadih in inhibiting the growth of bacteria Aggregatibacter actinomycetemcomitans. Material and Method: This was a true experimental study with post-test and control group design. This study was conducted on 36 samples of 2 treatment groups, the test group with probiotic Pediococcus pentosacesus isolated from dadih and the negative control group with sterile aquadest. The antibacterial effect was tested using the Kirby-Bauer disk diffusion method and calculated by measuring the zone of inhibition on MHA around paper disk using a sliding caliper with 0.5 mm accuracy. Result: The result of bivariate analysis using Independent t-test was p=0.00 where p < 0.05 means that there is a significant difference between the tested group and negative control group. Conclusion: Probiotic Pediococcus pentosaceus isolated from dadih are able to inhibit the growth of Aggregatibacter actinomycetemcomitans.

Keywords: aggregatibacter actinomycetemcomitans, antibacterial activities, periodontitis, probiotic Pediococcus pentosaceus

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248 Preliminary Studies on the Potentials of Bambara nut (Voandzeia substerranea) and Pigeon pea (Cajanus cajan) as Imitation Milk

Authors: Onuoha Gideon

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The preliminary studies on the potentials of Bambara nut and pigeon pea as imitation milk were investigated. Bambara nut and Pigeon pea milk were produced from two separate unit operations; Bambara nut seed was cooked, dehulled, milled and strained to milk (BCM) and another batch was toasted at moderate temperature, dehulled, milled and strained to milk (BTM). Pigeon pea seed was cooked, dehulled, milled and strained to milk (PCM) and another batch was toasted at moderate temperature, dehulled, milled and strained to milk (PTM). The result of the proximate analysis on the milk samples on wet basis showed that the protein content ranged from 28.56 – 26.77, the crude fibre ranged from 6.28 – 1.85, the ash content ranged from 5.22 – 1.17, the fat content ranged from 2.71 – 1.12, the moisture content ranged from 95.93 – 93.83, the carbohydrate content ranged from 67.62 – 58.83. The functional analysis on the milk samples showed that emulsification capacity ranged from 43.21 – 38.66, emulsion stability ranged from 34.10 – 25.00, the specific gravity ranged from 997.50 – 945.00, the foaming capacity ranged from 3,500 to 2,250, the measurement of viscosity ranged from 0.017 – 0.007, the pH range from 5.55 – 5.25, the measurement of dispersibility range from 11.00 – 7.00, the total soluble solid ranged from 4.00 to 1.75, the total titratable acidity ranged from 0.314 – 0.328. The sensory evaluation report showed that in terms of flavor, sample BCM and PCM value were significantly different from sample BTM and PTM. In terms of colour, sample BCM showed a significant difference from samples BTM, PCM and PTM. In term of texture, sample BCM was significantly different from samples BTM, PCM and PTM. The general acceptability shows that sample BCM was significantly different from other the samples and was the most accepted. The microbial analysis indicated that the microbial load increases with time. Bacterial count ranged from 1.3 x 105 – 1.20 x 106 to 1.6 x 105 – 1.06 x 106, fungal count ranged from 4.0 x 105 – 8.0 x 105 to 4.0 x 105 – 7.0 x 105. The studies showed that BCM was the most preferred.

Keywords: imitation milk, Bambara nut, Pigeon pea, proximate composition

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247 In silico Subtractive Genomics Approach for Identification of Strain-Specific Putative Drug Targets among Hypothetical Proteins of Drug-Resistant Klebsiella pneumoniae Strain 825795-1

Authors: Umairah Natasya Binti Mohd Omeershffudin, Suresh Kumar

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Klebsiella pneumoniae, a Gram-negative enteric bacterium that causes nosocomial and urinary tract infections. Particular concern is the global emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae. Characterization of antibiotic resistance determinants at the genomic level plays a critical role in understanding, and potentially controlling, the spread of multidrug-resistant (MDR) pathogens. In this study, drug-resistant Klebsiella pneumoniae strain 825795-1 was investigated with extensive computational approaches aimed at identifying novel drug targets among hypothetical proteins. We have analyzed 1099 hypothetical proteins available in genome. We have used in-silico genome subtraction methodology to design potential and pathogen-specific drug targets against Klebsiella pneumoniae. We employed bioinformatics tools to subtract the strain-specific paralogous and host-specific homologous sequences from the bacterial proteome. The sorted 645 proteins were further refined to identify the essential genes in the pathogenic bacterium using the database of essential genes (DEG). We found 135 unique essential proteins in the target proteome that could be utilized as novel targets to design newer drugs. Further, we identified 49 cytoplasmic protein as potential drug targets through sub-cellular localization prediction. Further, we investigated these proteins in the DrugBank databases, and 11 of the unique essential proteins showed druggability according to the FDA approved drug bank databases with diverse broad-spectrum property. The results of this study will facilitate discovery of new drugs against Klebsiella pneumoniae.

Keywords: pneumonia, drug target, hypothetical protein, subtractive genomics

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246 Advanced Biosensor Characterization of Phage-Mediated Lysis in Real-Time and under Native Conditions

Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka

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Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.

Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68

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245 The Culex Pipiens Niche: Assessment with Climatic and Physiographic Variables via a Geographic Information System

Authors: Maria C. Proença, Maria T. Rebelo, Marília Antunes, Maria J. Alves, Hugo Osório, Sofia Cunha, João Casaca

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Using a geographic information system (GIS), the relations between a georeferenced data set of Culex pipiens sl. mosquitoes collected in Portugal mainland during seven years (2006-2012) and meteorological and physiographic parameters such as: air relative humidity, air temperature (minima, maxima and mean daily temperatures), daily total rainfall, altitude, land use/land cover and proximity to water bodies are evaluated. Focus is on the mosquito females; the characterization of its habitat is the key for the planning of chirurgical non-aggressive prophylactic countermeasures to avoid ambient degradation. The GIS allow for the spatial determination of the zones were the mosquito mean captures has been above average; using the meteorological values at these coordinates, the limits of each parameter are identified/computed. The meteorological parameters measured at the net of weather stations all over the country are averaged by month and interpolated to produce raster maps that can be segmented according to the thresholds obtained for each parameter. The intersection of the maps obtained for each month show the evolution of the area favorable to the species through the mosquito season, which is from May to October at these latitudes. In parallel, mean and above average captures were related to the physiographic parameters. Three levels of risk could be identified for each parameter, using above average captures as an index. The results were applied to the suitability meteorological maps of each month. The Culex pipiens critical niche is delimited, reflecting the critical areas and the level of risk for transmission of the pathogens to which they are competent vectors (West Nile virus, iridoviruses, rheoviruses and parvoviruses).

Keywords: Culex pipiens, ecological niche, risk assessment, risk management

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244 Beneficiation of Pulp and Paper Mill Sludge for the Generation of Single Cell Protein for Fish Farming

Authors: Lucretia Ramnath

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Fishmeal is extensively used for fish farming but is an expensive fish feed ingredient. A cheaper alternate to fishmeal is single cell protein (SCP) which can be cultivated on fermentable sugars recovered from organic waste streams such as pulp and paper mill sludge (PPMS). PPMS has a high cellulose content, thus is suitable for glucose recovery through enzymatic hydrolysis but is hampered by lignin and ash. To render PPMS amenable for enzymatic hydrolysis, the PPMS waspre-treated to produce a glucose-rich hydrolysate which served as a feed stock for the production of fungal SCP. The PPMS used in this study had the following composition: 72.77% carbohydrates, 8.6% lignin, and 18.63% ash. The pre-treatments had no significant effect on lignin composition but had a substantial effect on carbohydrate and ash content. Enzymatic hydrolysis of screened PPMS was previously optimized through response surface methodology (RSM) and 2-factorial design. The optimized protocol resulted in a hydrolysate containing 46.1 g/L of glucose, of which 86% was recovered after downstream processing by passing through a 100-mesh sieve (38 µm pore size). Vogel’s medium supplemented with 10 g/L hydrolysate successfully supported the growth of Fusarium venenatum, conducted using standard growth conditions; pH 6, 200 rpm, 2.88 g/L ammonium phosphate, 25°C. A maximum F. venenatum biomass of 45 g/L was produced with a yield coefficient of 4.67. Pulp and paper mill sludge hydrolysate contained approximately five times more glucose than what was needed for SCP production and served as a suitable carbon source. We have shown that PPMS can be successfully beneficiated for SCP production.

Keywords: pulp and paper waste, fungi, single cell protein, hydrolysate

Procedia PDF Downloads 168