Search results for: protoplast isolation

Authors: Suman Chaudhary, Rinku Dhanker, Tanvi, Sneh Goyal

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Sulphur oxidizing bacteria (SOB) have marked their significant role in perspectives of maintaining healthy environment as researchers from all over the world tested and apply these in waste water treatment plants, bioleaching of heavy metals, deterioration of bridge structures, concrete and for bioremediation purposes, etc. Also, these SOB are well adapted in all kinds of environment ranging from normal soil, water habitats to extreme natural sources like geothermal areas, volcanic eruptions, black shale and acid rock drainage (ARD). SOB have been isolated from low pH environment of anthropogenic origin like acid mine drainage (AMD) and bioleaching heaps, hence these can work efficiently in different environmental conditions. Besides having many applications in field of environment science, they may be proven to be very beneficial in area of agriculture as sulphur is the fourth major macronutrients required for the growth of plants. More amount of sulphur is needed by pulses and oilseed crops with respect to the cereal grains. Due to continuous use of land for overproduction of more demanding sulphur utilizing crops and without application of sulphur fertilizers, its concentration is decreasing day by day, and thus, sulphur deficiency is becoming a great problem as it affects the crop productivity and quality. Sulphur is generally found in soils in many forms which are unavailable for plants (cannot be use by plants) like elemental sulphur, thiosulphate which can be taken up by bacteria and converted into simpler forms usable by plants by undergoing a series of transformations. So, keeping the importance of sulphur in view for various soil types, oilseed crops and role of microorganisms in making them available to plants, we made an effort to isolate, optimize, and characterize SOB. Three potential strains of bacteria were isolated, namely SSF7, SSA21, and SSS6, showing sulphate production of concentration, i.e. 2.268, 3.102, and 2.785 mM, respectively. Also, these were optimized for various culture conditions like carbon, nitrogen source, pH, temperature, and incubation time, and characterization was also done.

Keywords: sulphur oxidizing bacteria, isolation, optimization, characterization, sulphate production

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Authors: Pelin Balcik Ercin

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Introduction: ZEB transcription factor family member ZEB2, has a role in epithelial to mesenchymal transition during development and metastasis. The altered circulating extracellular miRNAs expression is observed in diseases, and extracellular miRNAs have an important role in cancer cell microenvironment. In ChIP-Seq study, the expression of miR-2110 was found to be regulated by ZEB2. In this study, the effects of miR2110 on cell proliferation and migration of hepatocellular carcinoma (HCC) cells were examined. Material and Methods: SNU398 cells transfected with mimic miR2110 (20nM) (HMI0375, Sigma-Aldrich) and negative control miR (HMC0002, Sigma-Aldrich). MicroRNA isolation was accomplished with miRVANA isolation kit according to manufacturer instructions. cDNA synthesis was performed expression, respectively, and calibrated with Ct of controls. The real-time quantitative PCR (RT-qPCR) reaction was performed using the TaqMan Fast Advanced Master Mix (Thermo Sci.). Ct values of miR2110 were normalized to miR-186-5p and miR16-5p for the intracellular gene. Cell proliferation analysis was analyzed with the xCELLigence RTCA System. Wound healing assay was analyzed with the ImageJ program and relative fold change calculated. Results: The mimic-miR-2110 transfected SNU398 cells nearly nine-fold (log2) more miR-2110 expressed compared to negative control transfected cells. The mimic-miR-2110 transfected HCC cell proliferation significantly inhibited compared to the negative control cells. Furthermore, miR-2110-SNU398 cell migration capacity was relatively four-fold decreased compared to negative control-miR-SNU398 cells. Conclusion: Our results suggest the miR-2110 inhibited cell proliferation and also miR-2110 negatively affect cell migration compared to control groups in HCC cells. These data suggest the complexity of microRNA EMT transcription factors regulation. These initial results are pointed out the predictive biomarker capacity of miR-2110 in HCC.

Keywords: epithelial to mesenchymal transition, EMT, hepatocellular carcinoma cells, micro-RNA-2110, ZEB2

Procedia PDF Downloads 125

Authors: Shamas Ul Hassan, Nasir Mukhtar, Sajjad Ur Rehman, Asghar Ali Mian, Iftikhar Hussain, Muhammad Safdar Anjum

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The Mycoplasma gallisepticum (MG) is the major cause of economic looses in birds which is transmitted by free flying birds in the environment. These demands for improving the biosecurity measures at farm level including proper disposal of farm mortality and other wastes along with the inclusion of zoos and wild life parks in the MG surveillance programme. For the purpose of doing surveillance of MG in different pet, game and free flying birds a total of 12 samples each of peacocks, pheasants, ducks, pigeons, parrots, and house crows were included in the first ever study of its nature in Pakistan. During the study, the relevant samples along with recording clinical and postmortem findings were subjected to sero-prevalence, culture isolation and PCR system. Further PCR being more sensitive proves to be a better epidemiological tool. Seropositive findings revealed in peacocks, pheasants, ducks, pigeons, parrots, and crows were 66.7%, 58.3%, 41.7%, 41.7%, 16.7% and 16.7% respectively with some free flying birds giving ambiguous reactions. Whereas in the same order the culture/isolation positive results were recorded as 25%, 16.7%, 8.3%, 16.7%, 16.7%, and 25%. The samples were further confirmed on the basis of 732 bp product in PCR system. High rate of prevalence of MG in the pet, game and free flying birds regardless to their clinical findings demands to improve the biosecurity measures at the farm level with the minimum interaction of these birds with commercial poultry. Further the proper and timely disposal of all sorts of carcasses contaminated litter and wasted feed in such ways that the free flying birds are denied of picking up at those wastages. Moreover, MG surveillance system including the advances diagnostic techniques in wildlife parks and zoos be devised with proper timely preventive and therapeutic measures. The study proves that a variety of birds other then chicken either with or without clinical exhibitions carry MG organism which could be the potential source of infection for commercial poultry. The routine surveillance will be done to reduce the economic losses in poultry production.

Keywords: epidemiology, Mycoplasma gallisepticum (MG), free flying birds, surveillance, PCR

Procedia PDF Downloads 420

Authors: Inas Raafat, Azza El Bassiouny, Waldemar L. Olszewsky, Nagui E. Mikhail, Mona Nossier, Nora E. I. El-Bassiouni, Mona Zoheiry, Houda Abou Taleb, Noha Abd El-Aal, Ali Baioumy, Shimaa Attia

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Background: Orthotopic liver transplantation is an established treatment for patients with severe acute and end-stage chronic liver disease. The shortage of donor organs continues to be the rate-limiting factor for liver transplantation throughout the world. Hepatocyte transplantation is a promising treatment for several liver diseases and can, also, be used as a "bridge" to liver transplantation in cases of liver failure. Aim of the work: This study was designed to develop a highly efficient protocol for isolation and transplantation of hepatocytes in experimental Lewis rat model to provide satisfactory guidelines for future application on humans.Materials and Methods: Hepatocytes were isolated from the liver by double perfusion technique and bone marrow cells were isolated by centrifugation of shafts of tibia and femur of donor Lewis rats. Recipient rats were subjected to sub-lethal dose of irradiation 2 days before transplantation. In a laparotomy operation the spleen was injected by freshly isolated hepatocytes and bone marrow cells were injected intravenously. The animals were sacrificed 45 day latter and splenic sections were prepared and stained with H & E, PAS AFP and Prox1. Results: The data obtained from this study showed that the double perfusion technique is successful in separation of hepatocytes regarding cell number and viability. Also the method used for bone marrow cells separation gave excellent results regarding cell number and viability. Intrasplenic engraftment of hepatocytes and live tissue formation within the splenic tissue were found in 70% of cases. Hematoxylin and eosin stained splenic sections from 7 rats showed sheets and clusters of cells among the splenic tissues. Periodic Acid Schiff stained splenic sections from 7 rats showed clusters of hepatocytes with intensely stained pink cytoplasmic granules denoting the presence of glycogen. Splenic sections from 7 rats stained with anti-α-fetoprotein antibody showed brownish cytoplasmic staining of the hepatocytes denoting positive expression of AFP. Splenic sections from 7 rats stained with anti-Prox1 showed brownish nuclear staining of the hepatocytes denoting positive expression of Prox1 gene on these cells. Also, positive expression of Prox1 gene was detected on lymphocytes aggregations in the spleens. Conclusions: Isolation of liver cells by double perfusion technique using collagenase buffer is a reliable method that has a very satisfactory yield regarding cell number and viability. The intrasplenic route of transplantation of the freshly isolated liver cells in an immunocompromised model was found to give good results regarding cell engraftment and tissue formation. Further studies are needed to assess function of engrafted hepatocytes by measuring prothrombin time, serum albumin and bilirubin levels.

Keywords: Lewis rats, hepatocytes, BMCs, transplantation, AFP, Prox1

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Authors: Carol N. Flores-Fernández, Guadalupe Espilco, Cynthia Esquerre, Amparo I. Zavaleta

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Saline environments represent a valuable source of enzymes with novel properties and particular features for application in food, pharmaceutical and chemical industry. This study focuses on the isolation and screening of hydrolase-producing bacteria from Chilca salterns and the evaluation of their biotechnological potential. Soil samples were collected from Chilca salterns in Peru. For the isolation, medium containing 0.2 % of yeast extract, 5 % of NaCl and 10 % of the soil sample was used. After 72 h of incubation at 37 °C, serial dilutions were made up to 10−12 dilutions, spread on agar plates with 0.5 % of yeast extract and 5 % of NaCl, and incubated at 37 °C for 48 h. Screening of hydrolase-producing bacteria was carried out for cellulases, amylases, lipases, DNase, and proteases on specific media. Moreover, protease-producing bacteria were tested using protein extracted from the following legumes as substrate: Glycine max, Lupinus mutabilis, Pisum sativum, Erythrina edulis, Cicer arietinum, Phaseolus vulgaris and Vicia faba. A total of 16 strains were isolated from soil samples. On the screening media; 75, 44, 81 and 50 % were cellulase, amylase, DNase and protease producers, respectively. Also, 19 % of the isolates produced all the hydrolytic enzymes above mentioned. Lipase producers were not found. The 37 % and 12 % of the strains grew at 20 % and 30 % of salt concentration, respectively. In addition, 75 % of the strains grew at pH range between 5 and 10. From the total of protease-producing bacteria, 100 % hydrolyzed Glycine max, Lupinus mutabilis, and Pisum sativum protein, while 87 % hydrolyzed Erythrina edulis and Cicer arietinum protein. Finally, 75 % and 50 % of the strains hydrolyzed Phaseolus vulgaris and Vicia faba protein, respectively. Hydrolase-producing bacteria isolated from Chilca salterns in Peru grew at high salt concentrations and wide range of pH. In addition, protease-producing bacteria hydrolyzed protein from different sources such as leguminous. These enzymes have great biotechnological potential and could be used for different industrial processes and applications.

Keywords: bacteria, extracellular, hydrolases, Peru, salterns

Procedia PDF Downloads 208

Authors: Habiba Nahzat, Karen Crow, Lisa Manuel, Maria Huijbregts

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Background and Rationale: The World Health Organization (WHO) declared COVID-19 a pandemic on March 11, 2020. Shortly after, the Ontario provincial and Toronto municipal governments also released multiple directives that led to the mass closure of businesses both in the public and private sectors. Recent research has identified connections between Intimate Partner Violence (IPV) and COVID-19 related stressors - especially because of lockdown and social isolation measures. Psychological impacts of lengthy seclusion coupled with disconnection from extended family and diminished support services can take a toll on families at risk and may increase mental health issues and the prevalence of IPV. Research Question: Thus, the purpose of the study was to understand the perspective of the Violence Against Women (VAW) program staff on the impact of the COVID-19 pandemic; we especially wanted to understand staff views of restrictions on clients’ counseling experiences and the ability to access services in general. The study also aimed to examine VAW program staff experiences regarding remote work and explore how the pandemic restriction measures affected the ability of their program operations to support their clients and each other. Method: A cross-sectional, descriptive qualitative study was conducted with a purposive sample of 9 VAW program staff – eight VAW counselors and one VAW manager. Prior to data collection, program staff collaborated in the development of the study purpose, interview questions and methodology. Ethics approval was obtained from the sponsoring organization’s Research Ethics Board. In-depth individual interviews were conducted with study participants using a semi-structured interview questionnaire. Brief demographic information was also collected prior to the interview. Descriptive statistics were used to analyze quantitative data and qualitative data was analyzed by thematic content analysis. Results: Findings from this study indicate that the COVID-19 pandemic restrictions had an adverse impact on clients seeking VAW services based on VAW staff perspectives. Program staff reported a perceived increase in abuse among women, especially in emotional and financial abuse and experiences of isolation and trauma. Findings further highlight the challenges women experienced when trying to access services in general as well as counseling and legal services. This was perceived to be more prominent among newcomers and marginalized women. The study also revealed client and staff challenges when participating in virtual counseling, their innovations and clients’ creativity in accessing needed counseling and how staff over time adapted to providing virtual support during the pandemic. Conclusion and Next Steps: This study builds upon existing evidence on the impact of COVID-19 restrictions on VAW and may inform future research to better understand the association between the COVID-19 pandemic restrictions and VAW on a broader scale and to inform and support possible short-term and long-term changes in the client experience and counselling practice.

Keywords: COVID-19, pandemic, virtual, violence against women (VAW)

Procedia PDF Downloads 189

Authors: Waled A. Dawoud, Abdelazim M. Negm, Nasser M. Saleh

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Offshore piling generates underwater sound at level high enough to cause physical damage or hearing impairment to the marine mammals. Several methods can be used to mitigate the effect of underwater noise from offshore pile driving on marine mammals which can be divided into three main approaches. The first approach is to keep the mammal out of the high-risk area by using aversive sound waves produced by acoustic mitigation devices such as playing-back of mammal's natural predator vocalization, alarm or distress sounds, and anthropogenic sound. The second approach is to reduce the amount of underwater noise from pile driving using noise mitigation techniques such as bubble curtains, isolation casing, and hydro-sound dampers. The third approach is to eliminate the overlap of underwater waves by using prolonged construction process. To investigate the effectiveness of different noise mitigation methods; a pile driven with 235 kJ rated energy diesel hammer near Jeddah Coast, Kingdom of Saudi Arabia was used. Using empirical sound exposure model based on Red Sea characteristics and limits of National Oceanic and Atmospheric Administration; it was found that the aversive sound waves should extend to 1.8 km around the pile location. Bubble curtains can reduce the behavioral disturbance area up to 28%; temporary threshold shift up to 36%; permanent threshold shift up to 50%; and physical injury up to 70%. Isolation casing can reduce the behavioral disturbance range up to 12%; temporary threshold shift up to 21%; permanent threshold shift up to 29%; and physical injury up to 46%. Hydro-sound dampers efficiency depends mainly on the used technology and it can reduce the behavioral disturbance range from 10% to 33%; temporary threshold shift from 18% to 25%; permanent threshold shift from 32% to 50%; and physical injury from 46% to 60%. To prolong the construction process, it was found that the single pile construction, use of soft start, and keep time between two successive hammer strikes more than 3 seconds are the most effective techniques.

Keywords: offshore pile driving, sound propagation models, noise effects on marine mammals, Underwater noise mitigation

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Authors: Nitin Rajan, Kashif Shakeel, Shashank Tiwari, Shachan Sagar

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Background: - Aegle Marmelos (Bael) leaf extract is taken twice daily to treat ophthalmia, ulcers, and intestinal worms, among other ailments. Poultice made from bael leaf is used in the treatment of eye conditions. The leaf juice has a variety of therapeutic applications, with the most notable being the treatment of diabetes. Fenugreek is used to cure red spots around the eyes, as well as to soften the throat and chest and to give relief from coughing. The use of this plant in the form of infusion, powder, pomade, and decoction has been extremely popular in Iranian traditional medicine. The plant may be used to wash one's vaginal linings. This plant is used as an emollient in the lack of appetite, treatment of pellagra, and gastrointestinal problems, as well as a general tonic. Calendula officinalis leaves are used to treat varicose veins on the outside of the body by infusing them. In Europe, the leaves are diaphoretic and resolvent in nature, while the blooms are employed as an emmenagogue and antispasmodic stimulant in Canada and the United States. The flowers were decocted and served as a posset drink when smallpox and measles were common in England, and the fresh juice was used to treat jaundice. Objective: - This study is done to compare the physicochemical parameter of the alcoholic extract of the leaves of Aegle Marmelos, Calendula Officinalis, and Fenugreek. Materials and Methods: Extraction and Isolation of Aegle Marmelos, Calendula Officinalis, Fenugreek, were done. Preliminary phytochemical study for alkaloids, cardiac glycosides, flavonoids, glycosides, phenols, resins, saponins, steroids, tannins, terpenoids of the extract was done individual by using the standard procedure. Result: - The phytochemical screening of Aegle Marmelos, Calendula Officinalis, and Fenugreek shows the presence of alkaloids, carbohydrates, total phenolics, total flavonoids, tannins, saponins gum. Conclusion: - In this study, we have found that crude aqueous and organic solvent extracts of Aegle Marmelos, Calendula Officinalis, and Fenugreek leaves contain some important bioactive compounds and it justifies their use in the traditional medicines for the treatment of different diseases.

Keywords: Aegle Marmelos, Calendula Officinalis, Fenugreek, physiochemical parameter

Procedia PDF Downloads 154

Authors: Karol Rodriguez-Peña, Martha L. Macias-Rubalcava, Leticia Rocha-Zavaleta, Sergio Sanchez

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A bioassay-guided study for anti-cancer compounds from endophytes of the Mexican medicinal plant Amphipteryygium adstringens resulted in the isolation of a streptomycete capable of producing a group of compounds with high cytotoxic activity. Microorganisms from surface sterilized samples of various sections of the plant were isolated and all the actinomycetes found were evaluated for their potential to produce compounds with cytotoxic activity against cancer cell lines MCF7 (breast cancer) and HeLa (cervical cancer) as well as the non-tumoural cell line HaCaT (keratinocyte). The most active microorganism was picked for further evaluation. The identification of the microorganism was carried out by 16S rDNA gene sequencing, finding the closest proximity to Streptomyces scabrisporus, but with the additional characteristic that the strain isolated in this study was capable of producing colorful compounds never described for this species. Crude extracts of dichloromethane and ethyl acetate showed IC50 values of 0.29 and 0.96 μg/mL for MCF7, 0.51 and 1.98 μg/mL for HeLa and 0.96 and 2.7 μg/mL for HaCaT. Scaling the fermentation to 10 L in a bioreactor generated 1 g of total crude extract, which was fractionated by silica gel open column to yield 14 fractions. Nine of the fractions showed cytotoxic activity. Fraction 4 was chosen for subsequent purification because of its high activity against cancerous cell lines, lower activity against keratinocytes. HPLC-UV-MS/ESI was used for the evaluation of this fraction, finding at least 10 different compounds with high values of m/z (≈588). Purification of the compounds was carried out by preparative thin-layer chromatography. The prevalent compound was Steffimycin B, a molecule known for its antibiotic and cytotoxic activities and also for its low solubility in aqueous solutions. Along with steffimycin B, another five compounds belonging to the steffimycin family were isolated and at this moment their structures are being elucidated, some of which display better solubility in water: an attractive property for the pharmaceutical industry. As a conclusion to this study, the isolation of endophytes resulted in the discovery of a strain capable of producing compounds with high cytotoxic activity that need to be studied for their possible utilization.

Keywords: amphipterygium adstringens, cytotoxicity, streptomyces scabrisporus, steffimycin

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Authors: Seyedeh Banafsheh Bagheri Marzouni, Sona Rostampour Yasouri

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Salmonella is one of the most important common pathogens between humans and animals worldwide. Globally, the prevalence of the disease in humans is due to the consumption of food contaminated with animal-derived Salmonella. These foods include eggs, red meat, chicken, and milk. Contamination of chicken and its products with Salmonella may occur at any stage of the chicken processing chain. Salmonella infection is usually not fatal. However, its occurrence is considered dangerous in some individuals, such as infants, children, the elderly, pregnant women, or individuals with weakened immune systems. If Salmonella infection enters the bloodstream, the possibility of contamination of tissues throughout the body will arise. Therefore, determining the potential risk of Salmonella at various stages is essential from the perspective of consumers and public health. The aim of this study is to isolate and identify Salmonella from chicken samples distributed in the Tehran market using the Gold standard culture method and PCR techniques based on specific genes, invA and ent. During the years 2022-2023, sampling was performed using swabs from the liver and intestinal contents of distributed chickens in the Tehran province, with a total of 120 samples taken under aseptic conditions. The samples were initially enriched in buffered peptone water (BPW) for pre-enrichment overnight. Then, the samples were incubated in selective enrichment media, including TT broth and RVS medium, at temperatures of 37°C and 42°C, respectively, for 18 to 24 hours. Organisms that grew in the liquid medium and produced turbidity were transferred to selective media (XLD and BGA) and incubated overnight at 37°C for isolation. Suspicious Salmonella colonies were selected for DNA extraction, and PCR technique was performed using specific primers that targeted the invA and ent genes in Salmonella. The results indicated that 94 samples were Salmonella using the PCR technique. Of these, 71 samples were positive based on the invA gene, and 23 samples were positive based on the ent gene. Although the culture technique is the Gold standard, PCR is a faster and more accurate method. Rapid detection through PCR can enable the identification of Salmonella contamination in food items and the implementation of necessary measures for disease control and prevention.

Keywords: culture, PCR, salmonella spp, salmonella enteritidis

Procedia PDF Downloads 72

Authors: Priyanka Sharma, Ranjita Das, Mohan C. Kalita, Debajit Thakur

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Background: Actinomycetes are the richest source of novel bioactive secondary metabolites such as antibiotics, enzymes and other therapeutically useful metabolites with diverse biological activities. The present study aims at the antimicrobial potential and genetic diversity of culturable Actinomycetes isolated from protected forest ecosystems of Assam which includes Kaziranga National Park (26°30˝-26°45˝N and 93°08˝-93°36˝E), Pobitora Wildlife Sanctuary (26º12˝-26º16˝N and 91º58˝-92º05˝E) and Gibbon Wildlife Sanctuary (26˚40˝-26˚45˝N and 94˚20˝-94˚25˝E) which are located in the North-eastern part of India. Northeast India is a part of the Indo-Burma mega biodiversity hotspot and most of the protected forests of this region are still unexplored for the isolation of effective antibiotic-producing Actinomycetes. Thus, there is tremendous possibility that these virgin forests could be a potential storehouse of novel microorganisms, particularly Actinomycetes, exhibiting diverse biological properties. Methodology: Soil samples were collected from different ecological niches of the protected forest ecosystems of Assam and Actinomycetes were isolated by serial dilution spread plate technique using five selective isolation media. Preliminary screening of Actinomycetes for an antimicrobial activity was done by spot inoculation method and the secondary screening by disc diffusion method against several test pathogens, including multidrug resistant Staphylococcus aureus (MRSA). The strains were further screened for the presence of antibiotic synthetic genes such as type I polyketide synthases (PKS-I), type II polyketide synthases (PKS-II) and non-ribosomal peptide synthetases (NRPS) genes. Genetic diversity of the Actinomycetes producing antimicrobial metabolites was analyzed through 16S rDNA-RFLP using Hinf1 restriction endonuclease. Results: Based on the phenotypic characterization, a total of 172 morphologically distinct Actinomycetes were isolated and screened for antimicrobial activity by spot inoculation method on agar medium. Among the strains tested, 102 (59.3%) strains showed activity against Gram-positive bacteria, 98 (56.97%) against Gram-negative bacteria, 92 (53.48%) against Candida albicans MTCC 227 and 130 (75.58%) strains showed activity against at least one of the test pathogens. Twelve Actinomycetes exhibited broad spectrum antimicrobial activity in the secondary screening. The taxonomic identification of these twelve strains by 16S rDNA sequencing revealed that Streptomyces was found to be the predominant genus. The PKS-I, PKS-II and NRPS genes detection indicated diverse bioactive products of these twelve Actinomycetes. Genetic diversity by 16S rDNA-RFLP indicated that Streptomyces was the dominant genus amongst the antimicrobial metabolite producing Actinomycetes. Conclusion: These findings imply that Actinomycetes from the protected forest ecosystems of Assam, India, are a potential source of bioactive secondary metabolites. These areas are as yet poorly studied and represent diverse and largely unscreened ecosystem for the isolation of potent Actinomycetes producing antimicrobial secondary metabolites. Detailed characterization of the bioactive Actinomycetes as well as purification and structure elucidation of the bioactive compounds from the potent Actinomycetes is the subject of ongoing investigation. Thus, to exploit Actinomycetes from such unexplored forest ecosystems is a way to develop bioactive products.

Keywords: Actinomycetes, antimicrobial activity, forest ecosystems, RFLP

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Authors: Miguel Angel Calvo Salve

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Building enclosures and façades not only have an aesthetic component they must also ensure thermal comfort and improve the acoustics and air quality in buildings. The role of facades design, its assemblies, and construction are key in developing a greener future in architecture. This research and study focus on the design of a multi-leaves building envelope, with a self-supporting wall of granite slabs. The study will demonstrate the advantages of its use in compare with the hanging stone veneer in a vented cladding system. Using the Design of the School of Music and Theatre of the Atlantic Area in Spain as a case study where the multi-leaves enclosure system consists in a self-supported outer leaf of large granite slabs of 15cm. of thickness, a vent cavity with thermal isolation, a brick wall, and a series of internal layers. The methodology used were simulations and data collected in building. The advantages of the self-supporting wall of granite slabs in the outer leaf (15cm). compared with a hanging stone veneer in a vented cladding system can summarize the goals as follows: Using the stone in more natural way, by compression. The weight of the stone slabs goes directly to a strip-footing and don't overload the reinforced concrete structure of the building. The weight of the stone slabs provides an external aerial soundproofing, preventing the sound transmission to the structure. The thickness of the stone slabs is enough to provide the external waterproofing of the building envelope. The self-supporting system with minimum anchorages allows having a continuous and external thermal isolation without thermal bridges. The thickness of ashlars masonry provides a thermal inertia that balances the temperatures between day and night in the external thermal insulation layer. The absence of open joints gives the quality of a continuous envelope transmitting the sensations of the stone, the heaviness in the facade, the rhythm of the music and the sequence of the theatre. The main cost of stone due his bigger thickness is more than compensated with the reduction in assembly costs. Don´t need any substructure systems for hanging stone veneers.

Keywords: self-supporting wall, stone cladding systems, hanging veneer cladding systems, sustainability of facade systems

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Authors: Orathai Srithongtham, Ploypailin Mekathepakorn, Tossaphong Buraman, Pontida Moonpradap, Rungrueng Kitpati, Chulapon Kratet, Worayuth Nak-ai, Suwaree Charoenmukkayanan, Peeranuch Keawkanya

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The COVID-19 pandemic was effect to the health security of the Thai people. The PHEM principle was essential to the surveillance, prevention, and control of COVID-19. This study aimed to present the process of prevention and control of COVID-19 from February 29, 2021- April 30, 2022, and the factors and conditions influent the successful outcome. The study areas were three provinces. The target group was 37 people, composed of public health personnel. The data was collected in-depth, and group interviews followed the non-structure interview guide and were analyzed by content analysis. The components of COVID-19 prevention and control were found in the process of PHEM as follows; 1) Emergency Operation Center (EOC) with an incidence command system (ICS) from the district to provincial level and to propose the provincial measure, 2) Provincial Communicable Disease Committee (PCDC) to decide the provincial measure 3) The measure for surveillance, prevention, control, and treatment of COVID-19, and 4) outcomes and best practices for surveillance and control of COVID-19. The success factors of 4S and EC were as follows; Space: prepare the quarantine (HQ, LQ), Cohort Ward (CW), field hospital, and community isolation and home isolation to face with the patient and risky group, Staff network from various organization and group cover the community leader and Health Volunteer (HV), Stuff the management and sharing of the medical and non-medical equipment, System of Covid-19 respond were EOC, ICS, Joint Investigation Team (JIT) and Communicable Disease Control Unit (CDCU) for monitoring the real-time of surveillance and control of COVID-19 output, Environment management in hospital and the community with Infections Control (IC) principle, and Culture in term of social capital on “the relationship of Isan people” supported the patient provide the good care and support. The structure of PHEM, Isan’s Culture, and good preparation was a significant factor in the three provinces.

Keywords: public health, emergency management, covid-19, pandemic

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Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar

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Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.

Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus

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Authors: Ounis Hadj Mohamed, Ounis Abdelhafid

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In order to illustrate the effect of damping on the response of a base-isolated building, a parametric study is led, taking into account the progressive variation of the damping ratio (10% to 30%) under different types of seismic excitations (near and far field). A time history analysis is used to determine the response of the structure in terms of relative displacement and understory drift at various levels of the building. Thus, the results show that the efficiency of the isolator increases with the assumed damping ratio, provided that this latter is less or equal to 20%. Beyond this value, the isolator becomes less convenient. Furthermore, a strong deviation of energy capacity by the LRB (Lead Rubber Bearing) system is recorded.

Keywords: damping, base isolation, LRB, seismic excitation, hysteresis

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Authors: A. Sardi, J. Zbitou, A. Errkik, L. El Abdellaoui, A. Tajmouati, M. Latrach

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This paper presents analysis and design of a wideband Wilkinson power divider for wireless applications. The design is accomplished by transforming the lengths and impedances of the quarter wavelength sections of the conventional Wilkinson power divider into U-shaped sections. The designed power divider is simulated by using ADS Agilent technologies and CST microwave studio software. It is shown that the proposed power divider has simple topology and good performances in terms of insertion loss, port matching and isolation at all operating frequencies (1.8 GHz, 2.45 GHz and 3.55 GHz).

Keywords: ADS agilent technologies, CST microwave studio, microstrip, wideband, wilkinson power divider

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Authors: Yeasir Arafat Alve, Nazmun Nahar, Salma BeguM

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Introduction / Rationale: Caregivers of stroke survivors have experienced financial, emotional, physical and mental anxiety and have influence of family bonding and social customs, where 80% of caregivers were women and majority of the patients were cared for by immediate family members for example a spouse, son/daughter, son-in-law, daughter-in-law, siblings and they are significantly feel burden as a caregiver. In Bangladeshi context, there has a limitation of knowledge about the level of caregiver burden. This study could be suggested the health professional to focus on the care giving stress to provide a better support to them and also it will be advisable to provide equivalent services for caregivers and their families. Objectives: The study finds out the socio-demographic image of caregivers of stroke survivors in Bangladesh as well as discovers the level of burden of caregiver of stroke survivor in relation to general strain, isolation, disappointment, emotional involvement and environment. The study will find out the association between level of burden among caregivers and onset of stroke of survivors & duration of care giving. As well as to determine the association between level of burden among caregivers and caregiver’s age, gender, occupation and caregiver’s relationship with stroke survivors. Method / Approach: The study is a non experimental cross-sectional study design where 151 participants were selected through purposive comprehensive sampling. Data were selected from occupational therapy outdoor and stroke rehab unit, CRP (Savar & Mirpur) where using the Caregiver Burden Scale (a structured questionnaire) with face to face interview. Results: Most of the caregivers (78.8%) of stroke survivors faced moderate level of burden in general strain (37.7%), isolation (27.2%) but in case of disappointment (60.3%) feel higher burden and lower burden in emotional involvement (9.9%) and environment (0.7%). Caregiver burden level was significantly associated with caregivers’ age (P=0.006), sex (P=0.002), occupation (p= 0.04), relationship with stroke survivors (P=0.02), care giving duration (P=0.000), care giving hours (P=0.009), and onset of stroke (P=0.000) of stroke survivors. Conclusion: The study findings revealed that most of the caregivers faced moderate burden where no environmental burden for them, this is possibly in case of Bangladeshi culture where people hospitable. Through this study, it was also found that there is a possibility to have the higher burden. Finally, it is being also suggested that appropriate advice and support may preserve care giving which eventually enables the survivors to live a longer and more fulfilling life in the community.

Keywords: caregiver, level of caregiver burden, stroke survivor, stroke rehab unit

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Authors: Cem Çindaş, Serkan Şimşek

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In this paper, the study and design of a 3-dB 90° directional coupler operating in the S-band is proposed. The coupler employs symmetrical multi-section coupled lines designed in a stripline technique. Design is realized in AWR Design Environment and CST Microwave Studio. Using these two programs played a key role in attaining outcomes swiftly and precisely. The simulation results show that the coupler maintains amplitude consistency within ± 0.3 dB, isolation and reflection losses better than 16 dB, and phase difference between two output ports of 88º±0.6˚ in the 1.7 – 4.35 GHz range. This simulation results indicate an improvement is achieved in fractional bandwidth (FBW) performance around the center frequency of f0 = 3 GHz.

Keywords: coupled stripline, directional coupler, multi-section coupler, symmetrical coupler

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Authors: Ryan Maclellan

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EXO-200 has provided one of the most sensitive searches for neutrinoless double-beta decay utilizing 175 kg of enriched liquid xenon in an ultra-low background time projection chamber. This detector has demonstrated excellent energy resolution and background rejection capabilities. Using the first two years of data, EXO-200 has set a limit of 1.1x10^25 years at 90% C.L. on the neutrinoless double-beta decay half-life of Xe-136. The experiment has experienced a brief hiatus in data taking during a temporary shutdown of its host facility: the Waste Isolation Pilot Plant. EXO-200 expects to resume data taking in earnest this fall with upgraded detector electronics. Results from the analysis of EXO-200 data and an update on the current status of EXO-200 will be presented.

Keywords: double-beta, Majorana, neutrino, neutrinoless

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Authors: Nicolò Vaiana, Filip C. Filippou, Giorgio Serino

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In order to reduce numerical computations in the nonlinear dynamic analysis of seismically base-isolated structures, a Mixed Explicit-Implicit time integration Method (MEIM) has been proposed. Adopting the explicit conditionally stable central difference method to compute the nonlinear response of the base isolation system, and the implicit unconditionally stable Newmark’s constant average acceleration method to determine the superstructure linear response, the proposed MEIM, which is conditionally stable due to the use of the central difference method, allows to avoid the iterative procedure generally required by conventional monolithic solution approaches within each time step of the analysis. The main aim of this paper is to investigate the stability and computational efficiency of the MEIM when employed to perform the nonlinear time history analysis of base-isolated structures with sliding bearings. Indeed, in this case, the critical time step could become smaller than the one used to define accurately the earthquake excitation due to the very high initial stiffness values of such devices. The numerical results obtained from nonlinear dynamic analyses of a base-isolated structure with a friction pendulum bearing system, performed by using the proposed MEIM, are compared to those obtained adopting a conventional monolithic solution approach, i.e. the implicit unconditionally stable Newmark’s constant acceleration method employed in conjunction with the iterative pseudo-force procedure. According to the numerical results, in the presented numerical application, the MEIM does not have stability problems being the critical time step larger than the ground acceleration one despite of the high initial stiffness of the friction pendulum bearings. In addition, compared to the conventional monolithic solution approach, the proposed algorithm preserves its computational efficiency even when it is adopted to perform the nonlinear dynamic analysis using a smaller time step.

Keywords: base isolation, computational efficiency, mixed explicit-implicit method, partitioned solution approach, stability

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Authors: Placide Poba-Nzaou, Anicet Tchibozo, Malatsi Galani, Ali Etkkali, Erwin Halim

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Artificial intelligence (AI) is a general-purpose technology that is transforming many industries, working life and society by stimulating economic growth and innovation. Despite the huge potential of benefits to be generated, the adoption of AI varies from one organization to another, from one region to another, and from one industry to another, due in part to obstacles that can inhibit an organization or organizations located in a specific geographic region or operating in a specific industry from adopting AI technology. In this context, these obstacles and their implications for AI adoption from the perspective of configurational theory is important for at least three reasons: (1) understanding these obstacles is the first step in enabling policymakers and providers to make an informed decision in stimulating AI adoption (2) most studies have investigating obstacles or challenges of AI adoption in isolation with linear assumptions while configurational theory offers a holistic and multifaceted way of investigating the intricate interactions between perceived obstacles and barriers helping to assess their synergetic combination while holding assumptions of non-linearity leading to insights that would otherwise be out of the scope of studies investigating these obstacles in isolation. This study aims to pursue two objectives: (1) characterize organizations by uncovering the typical profiles of combinations of 15 internal and external obstacles that may prevent organizations from adopting AI technology, (2) assess the variation in terms of intensity of AI adoption associated with each configuration. We used data from a survey of AI adoption by organizations conducted throughout the EU27, Norway, Iceland and the UK (N=7549). Cluster analysis and discriminant analysis help uncover configurations of organizations based on the 15 obstacles, including eight external and seven internal. Second, we compared the clusters according to AI adoption intensity using an analysis of variance (ANOVA) and a Tamhane T2 post hoc test. The study uncovers three strongly separated clusters of organizations based on perceived obstacles to AI adoption. The clusters are labeled according to their magnitude of perceived obstacles to AI adoption: (1) Cluster I – High Level of perceived obstacles (N = 2449, 32.4%)(2) Cluster II – Low Level of perceived obstacles (N =1879, 24.9%) (3) Cluster III – Moderate Level of perceived obstacles (N =3221, 42.7%). The proposed taxonomy goes beyond the normative understanding of perceived obstacles to AI adoption and associated implications: it provides a well-structured and parsimonious lens that is useful for policymakers, AI technology providers, and researchers. Surprisingly, the ANOVAs revealed a “high level of perceived obstacles” cluster associated with a significantly high intensity of AI adoption.

Keywords: Artificial intelligence (AI), obstacles, adoption, taxonomy.

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Authors: Kira Rysakova, Vitaly Novikov

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An improved method of obtaining enzymatic collagen hydrolysate from the tissues of marine hydrobionts is proposed, which allows to obtain hydrolysate without pre-isolation of pure collagen. The method can be used to isolate enzymatic collagen hydrolysate from the waste of industrial processing of Red King crab and non-traditional objects - marine holothurias. Comparative analysis of collagen hydrolysates has shown the possibility of their use in a number of nutrient media, but this requires additional optimization of their composition and biological tests on wide sets of test strains of microorganisms.

Keywords: collagen hydrolysate, marine hydrobionts, red king crab, marine holothurias, enzymes, exclusive HPLC

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Authors: Xin Li, Zhenxue Jiang, Zhuo Li

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Generally, shale gas is trapped within shale systems with low porosity and ultralow permeability as free and adsorbing states. Its production is controlled by properties, in terms of occurrence phases, gas contents, and percolation characteristics. These properties are all influenced by porous features. In this paper, porosity differences of marine shales were explored between Lower Cambrian shale and Lower Silurian shale of Sichuan Basin, South China. Both the two shales were marine shales with abundant oil-prone kerogen and rich siliceous minerals. Whereas Lower Cambrian shale (3.56% Ro) possessed a higher thermal degree than that of Lower Silurian shale (2.31% Ro). Samples were measured by a combination of organic-chemistry geology measurement, organic matter (OM) isolation, X-ray diffraction (XRD), N2 adsorption, and focused ion beam milling and scanning electron microscopy (FIB-SEM). Lower Cambrian shale presented relatively low pore properties, with averaging 0.008ml/g pore volume (PV), averaging 7.99m²/g pore surface area (PSA) and averaging 5.94nm average pore diameter (APD). Lower Silurian shale showed as relatively high pore properties, with averaging 0.015ml/g PV, averaging 10.53m²/g PSA and averaging 18.60nm APD. Additionally, fractal analysis indicated that the two shales presented discrepant pore morphologies, mainly caused by differences in the combination of pore types between the two shales. More specifically, OM-hosted pores with pin-hole shape and dissolved pores with dead-end openings were the main types in Lower Cambrian shale, while OM-hosted pore with a cellular structure was the main type in Lower Silurian shale. Moreover, porous characteristics of isolated OM suggested that OM of Lower Silurian shale was more capable than that of Lower Cambrian shale in the aspect of pore contribution. PV of isolated OM in Lower Silurian shale was almost 6.6 times higher than that in Lower Cambrian shale, and PSA of isolated OM in Lower Silurian shale was almost 4.3 times higher than that in Lower Cambrian shale. However, no apparent differences existed among samples with various matrix compositions. At late diagenetic or metamorphic epoch, extensive diagenesis overprints the effects of minerals on pore properties and OM plays the dominant role in pore developments. Hence, differences of porous features between the two marine shales highlight the effect of diagenetic degree on OM-hosted pore development. Consequently, distinctive pore characteristics may be caused by the different degrees of diagenetic evolution, even with similar matrix basics.

Keywords: marine shale, lower Cambrian, lower Silurian, om isolation, pore properties, om-hosted pore

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Authors: Somia Bouzid, Messaoud Ramdani

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The success of any diagnosis strategy critically depends on the sensors measuring process variables. This paper presents a detection and diagnosis sensor faults method based on a Bottleneck Neural Network (BNN). The BNN approach is used as a statistical process control tool for drinking water distribution (DWD) systems to detect and isolate the sensor faults. Variable reconstruction approach is very useful for sensor fault isolation, this method is validated in simulation on a nonlinear system: actual drinking water distribution system. Several results are presented.

Keywords: fault detection, localization, PCA, NLPCA, auto-associative neural network

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Authors: Preeti Panthari, Harsha Kharkwal

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Myrica nagi belongs to Myricaceae family. It is known for its therapeutic use since ancient times. The leaves were extracted with methanol and further fractioned with different solvents with increasing polarity. The n-butanol fraction of methanol extract was passed through celite, on separation through silica gel column chromatography yielded ten fractions. For the first time we report isolation of Caffeic acid from n-butanol fraction of Myrica nagi leaves in Chloroform: methanol (70:30) fraction. The mobile phase used for analysis in HPLC was Methanol: water (60:40) at the flow rate of 1 ml/min at wavelength of 280 nm. The retention time was 2.66 mins.

Keywords: Myrica nagi, column chromatography, retention time, caffeic acid

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Authors: Vasileios Gerakis, Fontounasios Christos, Alkis Hatzopoulos

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A study of the impact of metal guard rings on the coupling between two square metal pads is presented. The structure is designed over a bulk silicon substrate with epitaxial layer, so the coupling through the substrate is also involved. A lightly doped profile is adopted and is simulated by means of an electromagnetic simulator for various pad distances and different metal layers, assuming a 65 nm bulk CMOS technology. The impact of various guard ring design (geometrical) parameters is examined. Furthermore, the increase of isolation (resulting in reduction of the noise coupling) between the pads by cutting the ring, or by using multiple rings, is also analyzed. S parameters are used to compare the various structures.

Keywords: guard rings, metal pad coupling, millimeter wave frequencies, substrate noise,

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Authors: Somayeh Akrami, Vali-Allah Mozaffarian, Habib Onsori

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The studied region in this research, watershed Kuhkamar river, is about 112.66 square kilometers, it is located between 45º 48' 9" to 45º 2' 20" N and 38º 34' 15" to 38º 40' 28" E. The gained results of the studies on flora combinations, proved 287 plant species in 190 genera and 51 families. Asteracea with 49 and Lamiaceae with 27 plant species are the major plant families. Among collected species one interesting plant was found and determined as a new record Anemone narcissiflora L. for flora of Iran. This plant is known as a complex species that shows intraspecific speciation and is classified into about 12 subspecies and 10 varieties in world. To identify the infraspecies taxons of this species, in addition to morphological characteristics, the use of appropriate molecular markers for the better isolation of the individuals were needed.

Keywords: Anemone narcissiflora, floristic Study, kuhkamar, molecular marker

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Authors: Yiwei Li, Mingyu Gao

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Integrating hybrid memories with heterogeneous processors could leverage heterogeneity in both compute and memory domains for better system efficiency. To ensure performance isolation, we introduce Hydrogen, a hardware architecture to optimize the allocation of hybrid memory resources to heterogeneous CPU-GPU systems. Hydrogen supports efficient capacity and bandwidth partitioning between CPUs and GPUs in both memory tiers. We propose decoupled memory channel mapping and token-based data migration throttling to enable flexible partitioning. We also support epoch-based online search for optimized configurations and lightweight reconfiguration with reduced data movements. Hydrogen significantly outperforms existing designs by 1.21x on average and up to 1.31x.

Keywords: hybrid memory, heterogeneous systems, dram cache, graphics processing units

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Authors: A. Zh. Aupova, A. Ulankyzy, A. Sarsenova, A. Kussayin, Sh. Turarbek, N. Moldagulova, A. Kurmanbayev

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One of the methods of organic waste bioconversion into environmentally-friendly fertilizer is composting. Microorganisms that produce hydrolytic enzymes play a significant role in accelerating the process of organic waste composting. We studied the enzymatic potential (amylase, protease, cellulase, lipase, urease activity) of bacteria isolated from the sewage sludge of Nur-Sultan, Rudny, and Fort-Shevchenko cities, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha for processing organic waste and identifying active strains. Microorganism isolation was carried out by the cultures enrichment method on liquid nutrient media, followed by inoculating on different solid media to isolate individual colonies. As a result, sixty-one microorganisms were isolated, three of which were thermophiles (DS1, DS2, and DS3). The highest number of isolates, twenty-one and eighteen, were isolated from sewage sludge of Nur-Sultan and Rudny cities, respectively. Ten isolates were isolated from the wastewater of the sewage treatment plant in Fort-Shevchenko. From the dacha soil of Nur-Sultan city and freshly cut grass - 9 and 5 isolates were revealed, respectively. The lipolytic, proteolytic, amylolytic, cellulolytic, ureolytic, and oil-oxidizing activities of isolates were studied. According to the results of experiments, starch hydrolysis (amylolytic activity) was found in 2 isolates - CB2/2, and CB2/1. Three isolates - CB2, CB2/1, and CB1/1 were selected for the highest ability to break down casein. Among isolated 61 bacterial cultures, three isolates could break down fats - CB3, CBG1/1, and IL3. Seven strains had cellulolytic activity - DS1, DS2, IL3, IL5, P2, P5, and P3. Six isolates rapidly decomposed urea. Isolate P1 could break down casein and cellulose. Isolate DS3 was a thermophile and had cellulolytic activity. Thus, based on the conducted studies, 15 isolates were selected as a potential for sewage sludge composting - CB2, CB3, CB1/1, CB2/2, CBG1/1, CB2/1, DS1, DS2, DS3, IL3, IL5, P1, P2, P5, P3. Selected strains were identified on a mass spectrometer (Maldi-TOF). The isolate - CB 3 was referred to the genus Rhodococcus rhodochrous; two isolates CB2 and CB1 / 1 - to Bacillus cereus, CB 2/2 - to Cryseobacterium arachidis, CBG 1/1 - to Pseudoxanthomonas sp., CB2/1 - to Bacillus megaterium, DS1 - to Pediococcus acidilactici, DS2 - to Paenibacillus residui, DS3 - to Brevibacillus invocatus, three strains IL3, P5, P3 - to Enterobacter cloacae, two strains IL5, P2 - to Ochrobactrum intermedium, and P1 - Bacillus lichenoformis. Hence, 60 isolates were isolated from the wastewater of the cities of Nur-Sultan, Rudny, Fort-Shevchenko, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha. Based on the highest enzymatic activity, 15 active isolates were selected and identified. These strains may become the candidates for bio preparation for sewage sludge processing.

Keywords: sewage sludge, composting, bacteria, enzymatic activity

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

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An economically important disease of wheat in North Western region of India is Karnal Bunt caused by smut fungus Tilletia indica. This fungal pathogen spreads by air, soil and seed borne sporodia at the time of flowering, which ultimately leads to partial bunting of wheat kernels with fishy odor and taste to wheat flour. It has very serious effects due to quarantine measures which have to be applied for grain exports. Chemical fungicides such as mercurial compounds and Propiconazole applied to the control of Karnal bunt have been only partially successful. Considering the harmful effects of chemical fungicides on man as well as environment, many countries are developing biological control as the superior substitute to chemical control. Repeated use of fungicides can be responsible for the development of resistance in fungal pathogens against certain chemical compounds. The present investigation is based on the isolation and evaluation of antifungal properties of some isolated (from natural manure) and commercial bacterial strains against Tilletia indica. Total 23 bacterial isolates were obtained and antagonistic activity of all isolates and commercial bacterial strains (Bacillus subtilis MTCC8601, Bacillus pumilus MTCC 8743, Pseudomonas aeruginosa) were tested against T. indica by dual culture plate assay (pour plate and streak plate). Test for the production of antifungal volatile organic compounds (VOCs) by antagonistic bacteria was done by sealed plate method. Amongst all s1, s3, s5, and B. subtilis showed more than 80% inhibition. Production of extracellular hydrolytic enzymes such as protease, beta 1, 4 glucanase, HCN and ammonia was studied for confirmation of antifungal activity. s1, s3, s5 and B. subtilis were found to be the best for protease activity and s5 and B. subtilis for beta 1, 4 glucanase activity. Bacillus subtilis was significantly effective for HCN whereas s3, s5 and Bacillus subtilis for ammonia production. Isolates were identified as Pseudomonas aeruginosa (s1) and B. licheniformis (s3, s5) by various biochemical assays and confirmed by16s rRNA sequencing. Use of microorganisms or their secretions as biocontrol agents to avoid plant diseases is ecologically safe and may offer long term of protection to crop. The above study reports the promising effects of these strains in better pathogen free crop production and quality maintenance as well as prevention of the excessive use of synthetic fungicides.

Keywords: antagonistic, antifungal, biocontrol, Karnal bunt

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