Search results for: indigenous bacteria

Authors: Latifat Afolake Ogunfolabo, Hakeem Babafemi Ogunfolabo

Abstract:

The gastrointestinal tract is densely populated with micro-organism which closely and intensively interacts with the host and ingested feed. Food borne infections are some of the major international challenges that lead to high mortality and also, antimicrobial resistance, which has been classified as a serious threat by World Health Organization. Samples of slaughtered cattle and goats intestines were collected and standard culture methods were used for bacteria isolation and identification. Minimum inhibitory concentration of commonly used antibiotic using modification of the disk diffusion method was carried out on isolates. The samples cultured were all positive to Pseudomonas aeruginosa (95% and 90%), Escherichia coli (85%), Salmonella typhi (70% and 60%), Staphylococcus aureus (75%and 100%), Micrococcus luteus (55% and35%), Bacillus macerans (60% and 5%), Bacillus cereus (25% and 20%), Clostridium perfringens (20% and 5%), Micrococcus varians (20% and 5%), Bacillus subtilis (25% and 5%), Streptococcus faecalis (40% and 25%) and Streptococcus faecium (15% and 10%) in goat and cattle respectively. Also, Proteus mirabilis (40%), Micrococcus luteus (35%), Proteus vulgaris (30%), Klebsiella aerogenes(15%) were isolated from cattle. The total coliform (13.55 x10⁵cfu/gm ± 1.77) and (20.30 x10⁵cfu/gm ± 1.27) counts were significantly higher than the total bacteria count (8.3 x10⁵cfu/gm ± 1.41) and (16.60 x10⁵cfu/gm ±0.49) for goat and cattle respectively. Selected Bacteria count of isolates showed that Staphylococcus aureus had the highest significant value (6.9 x10⁵cfu/gm ± 0.57) and (16.80 x10⁵cfu/gm ± 0.57) Escherichia coli (4.60 x10⁵cfu/gm ± 0.42) and (7.05 x10⁵cfu/gm ± 0.64) while the lowest significant value was obtained in Salmonella/Shigella (1.7 x10⁵cfu/gm ± 0.00) and (1.5 x10⁵cfu/gm ± 0.00) for goat and cattle respectively. Susceptibility of bacteria isolated from slaughtered goat and cattle intestine to commonly used antibiotics showed that the highest statistical significant value for zone of inhibition for goat was obtained for Ciprofloxacin (30.00 ± 2.25, 23.75 ± 2.49, 17.17 ± 1.40) followed by Augmentin (28.33 ± 1.22, 21. 83 ± 2.44, 16.67 ± 1.49), Erythromycin (27.75 ±1.48, 20.25 ± 1.29, 16.67 ± 1.26) while the lowest values were obtained for Ofloxacin (27.17 ± 1.89, 21.42 ± 2.19, 16.83 ± 1.26) respectively and values obtained for cattle are Ciprofloxacin (30.64 ± 1.6, 25.79 ± 1.76, 8.07 ± 11.49) followed by Augmentin (28.29 ± 1.33, 22.64 ± 1.82, 17.43 ± 1.55) Ofloxacin (26.57 ± 2.02, 20.79 ± 2.75, 16.21 ± 1.19) while the lowest values were obtained for Erythromycin (26.64 ± 1.49, 20.29 ± 1.49, 16.29 ± 1.33) at different dilution factor (10⁻¹, 10⁻², 10⁻³) respectively. The isolates from goat and cattle were all susceptible to Augmentin at the three different dilution factors. Some goat isolates are intermediate to Ciprofloxacin and Erythromycin at 10⁻² and 10⁻³, while resistance to Ciprofloxacin at 10⁻³ dilution factor. Ciprofloxacin and Ofloxacin at the dilution factors of 10⁻³ and 10⁻¹ for some cattle isolate and resistance were observed for Ofloxacin and Erythromycin at dilution of 10⁻³. These results indicate the susceptibilities and the antimicrobial resistance to commonly used antibiotic.

Keywords: antibiotic susceptibility, bacteria, cattle, goat, identification

Procedia PDF Downloads 95

Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

Abstract:

Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover, this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

Procedia PDF Downloads 470

Authors: Ayuko Itsuki, Sachiyo Aburatani

Abstract:

Soil neutral sugar contents in Kasuga-yama Hill Primeval Forest, which is a World Heritage Site in Nara, Japan consisting of lowland laurel-leaved forest where natural conditions have been preserved for more than 1,000 years, were examined using the two-step hydrolysis to clarify the source of the neutral sugar and relations with the neutral sugar constituted the soil organic matter and the microbial biomass. Samples were selected from the soil (L, F, H and A horizons) surrounding laurel-leaved (BB-1) and Carpinus japonica (BB-2 and PW) trees for analysis. The neutral sugars were one factor of increasing the fungal and bacterial biomass in the laurel-leaved forest soil (BB-1). The more neutral sugar contents in the Cryptomeria japonica forest soil (PW) contributed to the growth of the bacteria and fungi than those of in the Cryptomeria japonica forest soil (BB-2). The neutral sugars had higher correlation with the numbers of bacteria and fungi counted by the dilution plate count method than by the direct microscopic count method. The numbers of fungi had higher correlation with those of bacteria by the dilution plate method.

Keywords: forest soil, neutral sugars, soil organic matter, two-step hydrolysis

Procedia PDF Downloads 238

Authors: G. Fouché, J. N. Eloff, K. Wellington

Abstract:

Acaricides are commonly used to control ticks but are toxic, harmful to the environment and too expensive to resource-limited farmers. Traditionally, many communities in South Africa rely on a wide range of indigenous practices to keep their livestock healthy. One of these health care practices includes the use of medicinal plants and this offers an alternative to conventional medicine. An investigation was conducted at the CSIR in South Africa, and selected indigenous plants used in communities were scientifically evaluated for the management of ticks in animals. 17 plants were selected from 239 plants used traditionally in South Africa. Two different organic extracts were prepared from the 17 samples, resulting in 34 plant samples. These were tested for efficacy against two tick species, namely Rhipicephalus microplus and Rhipicephalus turanicus. The plant extracts were also screened against Vero cells and most were found to have low cytotoxicity. This study has shown that there is potential for the development of botanicals as natural acaricides against ticks that are non-toxic and environmentally benign.

Keywords: South Africa, ticks, plant extracts, Rhipicephalus (Boophilus) microplus

Procedia PDF Downloads 196

Authors: Bisi-Johnson, Mary A., Hamzart A. Oyelade, Kehinde A. Adediran, Saheed A. Akinola

Abstract:

Geophagic and cosmetic clays are among potential nano-material which occur naturally and are of various forms. The use of these nano-clays is a common practice in both rural and urban areas mostly due to tradition and medicinal reasons. These naturally occurring materials can be valuable sources of nano-material by serving as nano-composites. The need to ascertain the safety of these materials is the motivation for this research. Physical Characterization based on the hue value and microbiological qualities of the nano-clays were carried out. The Microbial analysis of the clay samples showed considerable contamination with both bacteria and fungi with fungal contaminants taking the lead. This observation may not be unlikely due to the ability of fungi species to survive harsher growth conditions than bacteria. 'Atike pupa' showed no bacterial growth. The clay with the largest bacterial count was Calabash chalk (Igbanke), while that with the highest fungal count was 'Eko grey'. The most commonly isolated bacteria in this study were Clostridium spp. and Corynebacterium spp. while fungi included Aspergillus spp. These results are an indication of the need to subject these clay materials to treatments such as heating before consumption or topical usage thereby ascertaining their safety.

Keywords: nano-material, clay, microorganism, quality

Procedia PDF Downloads 356

Authors: Varsha Shinde, Belle Damodara Shenoy

Abstract:

Tarballs are crude oil remnants found in oceans after long term weathering process and are a global concern since several decades as potential marine pollutant. Being complicated in structure microbial remediation of tarballs in natural environment is a slow process. They are rich in high molecular weight alkanes and poly aromatic hydrocarbons which are resistant to microbial attack and other environmental factors, therefore remain in environment for long time. However, it has been found that many bacteria and fungi inhabit on tarballs for nutrients and shelter. Many of them are supposed to be oil degraders, while others are supposed to be getting benefited by byproducts formed during hydrocarbon metabolism. Thus tarballs are forming special interesting ecological niche of microbes. This work aimed to study diversity of bacteria and fungi from tarballs and to see their potential application in crude oil degradation. The samples of tarballs were collected from Betul beach of south Goa (India). Different methods were used to isolate culturable fraction of bacteria and fungi from it. Those were sequenced for 16S rRNA gene and ITS for molecular level identification. The 16S rRNA gene sequence analysis revealed the presence of 13 bacterial genera/clades (Alcanivorax, Brevibacterium, Bacillus, Cellulomonas, Enterobacter, Klebsiella, Marinobacter, Nitratireductor, Pantoea, Pseudomonas, Pseudoxanthomonas, Tistrella and Vibrio), while the ITS sequence analysis placed the fungi in 8 diverse genera/ clades (Aspergillus, Byssochlamys, Monascus, Paecilomyces, Penicillium, Scytalidium/ Xylogone, Talaromyces and Trichoderma). All bacterial isolates were screened for oil degradation capacity. Potential strains were subjected to crude oil degradation experiment for quantification. Results were analyzed by GC-MS-MS.

Keywords: bacteria, biodegradation, crude oil, diversity, fungi, tarballs

Procedia PDF Downloads 198

Authors: T. A. Rosy, M. R. I. Talukdar, N. S. Juyena, F. Y. Bari, M. N. Islam

Abstract:

The ovarian follicular dynamics, vaginal electrical resistance (VER) and progesterone (P4) and estrogen (E2) profiles were investigated during estrus cycle in four indigenous ewes. Daily VER values were recorded with heat detector. The follicles were observed and measured by trans-rectal ultrasonography. Blood was collected daily for hormonal profiles. Results showed a significant variation in VER values (P<0.05) at estrus in regards to ewes and cycles. The day difference between two successive lower values in VER waves ranged from 13-17 days which might indicate the estrus cycle in indigenous ewes. Trans-rectal ultrasonography of ovaries revealed the presence of two to four waves of follicular growth during the study period. Results also showed that follicular diameter was negatively correlated with VER values. Study of hormonal profiles by ELISA revealed a positive correlation between E2 concentration and development of follicle and negative correlation between P4 concentration and development of follicle. The concentrations of estradiol increased at the time of estrus and then fall down in a basal level. Development of follicular size was accompanied by an increase in the concentration of serum estradiol. Inversely, when follicles heed to ovulation concentration of progesterone starts to fall down and after ovulation it turns its way to the zenith and remains at this state until next ovulatory follicle comes to its maximum diameter. This study could help scientists to set up a manipulative reproductive technique for improving genetic values of sheep in Bangladesh.

Keywords: ovarian follicle, hormonal profile, sheep, ultrasonography, vaginal electrical resistance

Procedia PDF Downloads 240

Authors: Bhargavi G. Iyer, Ojaswi Bhagat

Abstract:

In recent years, there has been a lot of discussion about ethnic imbalance and diversity in the international context. Arbitration is now subject to the hegemony of a small number of people who are constantly reappointed. When a court system becomes exclusionary, the quality of adjudication suffers significantly. In such a framework, there is a misalignment between adjudicators' preconceived views and the interests of the parties, resulting in a biased view of the proceedings. The world is currently witnessing a slew of intellectual property battles around cultural appropriation. The term "cultural appropriation" refers to the industrial west's theft of indigenous culture, usually for fashion, aesthetic, or dramatic purposes. Selena Gomez exemplifies cultural appropriation by commercially using the “bindi,” which is sacred to Hinduism, as a fashion symbol. In another case, Victoria's Secret insulted indigenous peoples' genocide by stealing native Indian headdresses. In the case of yoga, a similar process can be witnessed, with Vedic philosophy being reduced to a type of physical practice. Such a viewpoint is problematic since indigenous groups have worked hard for generations to ensure the survival of their culture, and its appropriation by the western world for purely aesthetic and theatrical purposes is upsetting to those who practise such cultures. Because such conflicts involve numerous jurisdictions, they must be resolved through international arbitration. However, these conflicts are already being litigated, and the aggrieved parties, namely developing nations, do not believe it prudent to use the World Intellectual Property Organization's (WIPO) already established arbitration procedure. This practise, it is suggested in this study, is the outcome of Europe's exclusionary arbitral system, which fails to recognise the non-legal and non-commercial nature of indigenous culture issues. This research paper proposes a more comprehensive, inclusive approach that recognises the non-legal and non-commercial aspects of IP disputes involving cultural appropriation, which can only be achieved through an ethnically balanced arbitration structure. This paper also aspires to expound upon the benefits of arbitration and other means of alternative dispute resolution (ADR) in the context of disputes pertaining to cultural issues; positing that inclusivity is a solution to the existing discord between international practices and localised cultural points of dispute. This paper also hopes to explicate measures that will facilitate ensuring inclusion and ideal practices in the domain of arbitration law, particularly pertaining to cultural heritage and indigenous expression.

Keywords: arbitration law, cultural appropriation, dispute resolution, heritage, intellectual property

Procedia PDF Downloads 117

Authors: Sabah El-Sawalhi, Elie Fayad, Roula M. Abdel-Massih

Abstract:

Ilex paraguariensis (Yerba Mate) is a medium to large tree commonly consumed by South Americans. Its leaves and stems are associated with different biological activities. The purpose of this study was to evaluate the antibacterial activity of Yerba Mate against Gram-positive and Gram-negative bacterial strains and its action against some resistant bacteria with different resistance profiles. Yerba Mate aqueous extracts were prepared at 70°C for 2 hrs, and the microdilution method was used to determine the minimum inhibitory concentration (MIC). Gram-positive bacteria exhibited a stronger antibacterial activity (MIC ranged between 0.468 mg/mL and 15 mg/mL) than Gram-negative bacteria. Yerba Mate was also extracted with acetone: water (1:1) and then further sub-fractionated with hexane, chloroform, and ethyl acetate. MIC values against Staphylococcus aureus ranged from 0.78 to 2.5 mg/ml for the chloroform fraction, from 1.56 to 3.75 mg/ml for the ethyl acetate fraction, and 0.78 to 1.87 mg/ml for the water fraction. The water fraction also exhibited antibacterial activity against Salmonella species (MIC ranged from 1.56 mg/ml to 3.12 mg/ml). The water fraction exhibited the highest antibacterial activity among all the fractions obtained. More studies are needed to determine the molecule or molecules responsible for this activity.

Keywords: antibacterial activity, bacterial resistance, minimum inhibitory concentration, yerba mate

Procedia PDF Downloads 112

Authors: Thangaraj Arasakumar, Athar Ata, Palathurai Subramaniam Mohan

Abstract:

A series of novel 4-quinolone-3-carboxylic acid grafted spiropyrrolidines as new type of antibacterial agents were synthesized via multicomponent 1,3-dipolar cycloaddition reaction of an azomethine ylides with a newly prepared (E)-4-oxo-6-(3-phenyl-acryloyl)-1,4-dihydro-quinoline-3-carboxylic acids in high regioselectivity with good yields. The structure of cycloadduct characterized by FT IR, mass, 1H, 13C, 2D NMR techniques and elemental analysis. Structure and spectrometry of compound 8a has been investigated theoretically by using HF and DFT approach at B3LYP, M05-2x/6-31G* levels of theories. The optimized geometries and calculated vibrational frequencies are evaluated via comparison with experimental values. A good agreement is found between the measured and calculated values. The DFT studies support the molecular mechanism of this cycloaddition reaction and determine the molecular electrostatic potential and thermodynamic properties. Furthermore, the antibacterial activities of synthesized compounds were evaluated against Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and Gram-negative bacteria strains (Escherichia coli, Klebsiella pneumoniae). Among 21 compounds screened, 8f and 8p were found to be more active against tested bacteria.

Keywords: antibacterial activity, azomethine ylide, DFT calculation, spirooxindole

Procedia PDF Downloads 181

Authors: Te Kīpa Kēpa B. Morgan, Oliver W. Mcmillan, Dylan N. Taute, Tumanako N. Fa'aui

Abstract:

Epistemologies are ways of knowing. Indigenous Peoples are aware that they do not perceive and experience the world in the same way as others. So it is important when empowering Indigenous epistemologies, such as that of the New Zealand Māori, to also be able to represent a scientific understanding within the same analysis. A geothermal development assessment tool has been developed by adapting the Mauri Model Decision Making Framework. Mauri is a metric that is capable of representing the change in the life-supporting capacity of things and collections of things. The Mauri Model is a method of grouping mauri indicators as dimension averages in order to allow holistic assessment and also to conduct sensitivity analyses for the effect of worldview bias. R-shiny is the coding platform used for this Vision Mātauranga research which has created an expert decision support tool (DST) that combines a stakeholder assessment of worldview bias with an impact assessment of mauri-based indicators to determine the sustainability of proposed geothermal development. The initial intention was to develop guidelines for quantifying mātauranga Māori impacts related to geothermal resources. To do this, three typical scenarios were considered: a resource owner wishing to assess the potential for new geothermal development; another party wishing to assess the environmental and cultural impacts of the proposed development; an assessment that focuses on the holistic sustainability of the resource, including its surface features. Indicator sets and measurement thresholds were developed that are considered necessary considerations for each assessment context and these have been grouped to represent four mauri dimensions that mirror the four well-being criteria used for resource management in Aotearoa, New Zealand. Two case studies have been conducted to test the DST suitability for quantifying mātauranga Māori and other biophysical factors related to a geothermal system. This involved estimating mauri0meter values for physical features such as temperature, flow rate, frequency, colour, and developing indicators to also quantify qualitative observations about the geothermal system made by Māori. A retrospective analysis has then been conducted to verify different understandings of the geothermal system. The case studies found that the expert DST is useful for geothermal development assessment, especially where hapū (indigenous sub-tribal grouping) are conflicted regarding the benefits and disadvantages of their’ and others’ geothermal developments. These results have been supplemented with evaluations for the cumulative impacts of geothermal developments experienced by different parties using integration techniques applied to the time history curve of the expert DST worldview bias weighted plotted against the mauri0meter score. Cumulative impacts represent the change in resilience or potential of geothermal systems, which directly assists with the holistic interpretation of change from an Indigenous Peoples’ perspective.

Keywords: decision support tool, holistic geothermal assessment, indigenous knowledge, mauri model decision-making framework

Procedia PDF Downloads 159

Authors: Waad Al Saleemi, Badriya Al Adawi, Zaaima Al Jabri, Sahim Al Ghafri, Jalila Al Hadhramia

Abstract:

Objectives: Using standard lab methods, a positive blood culture requires a minimum of two days (two occasions of overnight incubation) to obtain a final identification (ID) and antimicrobial susceptibility results (AST) report. In this study, we aimed to evaluate the accuracy and precision of identification and antimicrobial susceptibility testing of an alternative method (direct method) that will reduce the turnaround time by 24 hours. This method involves the direct inoculation of positive blood culture broths into the Phoenix system using serum separation tubes (SST). Method: This prospective study included monomicrobial-positive blood cultures obtained from January 2022 to May 2023 in SQUH. Blood cultures containing a mixture of organisms, fungi, or anaerobic organisms were excluded from this study. The result of the new “direct method” under study was compared with the current “standard method” used in the lab. The accuracy and precision were evaluated for the ID and AST using Clinical and Laboratory Standards Institute (CLSI) recommendations. The categorical agreement, essential agreement, and the rates of very major errors (VME), major errors (ME), and minor errors (MIE) for both gram-negative and gram-positive bacteria were calculated. Passing criteria were set according to CLSI. Result: The results of ID and AST were available for a total of 158 isolates. Of 77 isolates of gram-negative bacteria, 71 (92%) were correctly identified at the species level. Of 70 isolates of gram-positive bacteria, 47(67%) isolates were correctly identified. For gram-negative bacteria, the essential agreement of the direct method was ≥92% when compared to the standard method, while the categorical agreement was ≥91% for all tested antibiotics. The precision of ID and AST were noted to be 100% for all tested isolates. For gram-positive bacteria, the essential agreement was >93%, while the categorical agreement was >92% for all tested antibiotics except moxifloxacin. Many antibiotics were noted to have an unacceptable higher rate of very major errors including penicillin, cotrimoxazole, clindamycin, ciprofloxacin, and moxifloxacin. However, no error was observed in the results of vancomycin, linezolid, and daptomycin. Conclusion: The direct method of ID and AST for positive blood cultures using SST is reliable for gram negative bacteria. It will significantly decrease the turnaround time and will facilitate antimicrobial stewardship.

Keywords: bloodstream infection, oman, direct ast, blood culture, rapid identification, antimicrobial susceptibility, phoenix, direct inoculation

Procedia PDF Downloads 25

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

Abstract:

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

Procedia PDF Downloads 316

Authors: Sameh Abu El-Soud, Zahra Zayed, Safwan Khedr, Adel M. Belal

Abstract:

The population increase in Egypt is urging for horizontal land development which became a demand to allow the benefit of different natural resources and expand from the narrow Nile valley. However, this development is facing challenges preventing land development and agriculture development. Desertification and moving sand dunes in the west sector of Egypt are considered the major obstacle that is blocking the ideal land use and development. In the proposed research, the sandy soil is treated biologically using Bacillus pasteurii bacteria as these bacteria have the ability to bond the sand partials to change its state of loose sand to cemented sand, which reduces the moving ability of the sand dunes. The procedure of implementing the Microbial Induced Carbonate Precipitation Technique (MICP) technique is examined, and the different factors affecting on this process such as the medium of bacteria sample preparation, the optical density (OD600), the reactant concentration, injection rates and intervals are highlighted. Based on the findings of the MICP treatment for sandy soil, conclusions and future recommendations are reached.

Keywords: soil stabilization, biological treatment, microbial induced carbonate precipitation (MICP), sand cementation

Procedia PDF Downloads 216

Authors: Mohammad Reza Haj Seyed Hadi, Fereshteh Ghanepasand, Mohammad Taghi Darzi

Abstract:

The main objective of this study was to determine the effects of Nitrogen fixing bacteria and manure application on the seed yield and yield components in black cumin (Nigella sativa L.). The experiment was carried out at the RAN Research Station in Firouzkouh in 2012. A 4×4 factorial experiment, arranged in a randomized complete blocks designed with three replications. The treatments consisted of 4 level of nitrogen fixing bacteria (control, Azotobacter, Azospirillum and Azotobacter + Azospirillum) and 4 level of manure (0, 2.5, 5 and 7.5 ton ha-1). The present results have shown that the highest height, 1000 seeds weight, seed number per follicle, follicle yield, seed yield and harvest index were obtained after using Azotobacter and Azospirillum, simultaneously. Manure application only affects on follicle yield and by 5ton manure ha-1 the highest follicle yield obtained. Results of this investigation showed that the maximum seed yield obtained when Aotobacter+Azospirillum inoculated with black cumin seeds and 5 ton manure ha-1 applied. Combined application of nitrogen fixing bacteria and manure can be helpful in developing of production and yield in Black cumin.

Keywords: azotobacter, azospirillum, black cumin, yield, yield components

Procedia PDF Downloads 426

Authors: Sanam Pudasaini, A. T. K. Perera, Ahmed Syed Shaheer Uddin, Sum Huan Ng, Chun Yang

Abstract:

Development of high-efficiency and environment friendly bacterial inactivation methods is of great importance for preventing waterborne diseases which are one of the leading causes of death in the world. Traditional bacterial inactivation methods (e.g., ultraviolet radiation and chlorination) have several limitations such as longer treatment time, formation of toxic byproducts, bacterial regrowth, etc. Recently, an electroporation-based inactivation method was introduced as a substitute. Here, an electroporation-based continuous flow microfluidic device equipped with an array of micropillars is developed, and the device achieved high bacterial inactivation performance ( > 99.9%) within a short exposure time ( < 1 s). More than 99.9% reduction of Escherichia coli bacteria was obtained for the flow rate of 1 mL/hr, and no regrowth of bacteria was observed. Images from scanning electron microscope confirmed the formation of electroporation-induced nano-pore within the cell membrane. Through numerical simulation, it has been shown that sufficiently large electric field strength (3 kV/cm), required for bacterial electroporation, were generated using PDMS micropillars for an applied voltage of 300 V. Further, in this method of inactivation, there is no involvement of chemicals and the formation of harmful by-products is also minimum.

Keywords: electroporation, high-efficiency, inactivation, microfluidics, micropillar

Procedia PDF Downloads 156

Authors: Irina E. Sukovataya, Oleg S. Sutormin, Valentina A. Kratasyuk

Abstract:

Effect of viscosity of media on kinetic parameters of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase was investigated with addition of organic solvents (glycerol and sucrose), because bioluminescent enzyme systems based on bacterial luciferases offer a unique and general tool for analysis of the many analytes and enzymes in the environment, research, and clinical laboratories and other fields. The possibility of stabilization and increase of activity of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase activity in vicious aqueous-organic mixtures have been shown.

Keywords: coupled enzyme system of bioluminescence bacteria NAD(P)H:FMN-oxidoreductase–luciferase, glycerol, stabilization of enzymes, sucrose

Procedia PDF Downloads 366

Authors: Muhammad Raqib Rasul, Tavga Sulaiman Rashid

Abstract:

Fertilization increases efficiency and obtains better quality of product recovery in agricultural activities. However, fertilizer consumption increased exponentially throughout the world, causing severe environmental problems. Biofertilizers can be a practical approach to minimize chemical fertilizer sources and ultimately develop soil fertility. This study was carried out to isolate, identify and characterize bacteria from medicinal plant (Rumex tuberosus L. and Verbascum sp.) rhizosphere for in vivo screening. 25 bacterial isolates were isolated and several biochemical tests were performed. Two isolates that were positive for most biochemical tests were chosen for the field experiment. The isolates were identified as Go1 Alcaligenes faecalis (Accession No. OP001725) and T11 (Bacillus sp.) based on the 16S rRNA sequence analysis that was compared with related bacteria in GenBank database using MEGA 6.1. For the field trial isolate GO1 and T11 (separately and mixed), NPK as a positive control was used. Both isolates increased plant height, chlorophyll content, number of tubers, and tuber’s weight. The results demonstrated that these two isolates of bacteria can potentially replace with chemical fertilizers for potato production.

Keywords: biofertilizer, Bacillus subtilis, Alcaligenes faecalis, potato tubers, in vivo screening

Procedia PDF Downloads 77

Authors: B. D. Ravindra, A. K. Tyagi, C. Kathirvelan

Abstract:

Some micronutrients in food (milk and meat), called ‘functional food components’ exert beneficial effects other than their routine nutrient function and conjugated linoleic acid (CLA) is an unsaturated fatty acid of ruminant origin, an example of this category. However, recently the fear of hypercholesterolemia due to saturated fats has led to the avoidance of dietary fat especially of animal origin despite its advantages such as lowering blood cholesterol, immuno-modulation and anticarcinogenic property due to the presence of CLA. The dietary increase of linoleic acid (LA) and linolenic acid (LNA) is one of the feeding strategies for increasing the CLA concentration in milk. Butyrivibrio fibrisolvens is the one potential rumen bacteria, which has high potential to isomerize LA to CLA. The study was conducted to screen the different oils for CLA production, selected based on their LA concentration. Butyrivibrio fibrisolvens culture (strain 49, MZ3, 30/10) were isolated from the rumen liquor of fistulated Buffalo (age ≈ 3 years; weight ≈ 250 kg) were used in in-vitro experiments, further work was carried out with three oils viz., sunflower, mustard and soybean oil at different concentration (0.05, 0.1, 0.15, 0.2, 0.25 and 0.3 g/L of media) to study the growth of bacteria and CLA production at different incubation period (0, 8, 12, 18, 24, 48, 72 h). In the present study, growth of the bacteria was decreased linearly with increase in concentration of three oils. However, highest decrease in growth was recorded at the concentration of 0.30 g of three oils per litre of the media. Highest CLA production was 51.96, 42.08 and 25.60 µg/ml at 0.25 g and it decreased to 48.19, 39.35 and 23.41 µg/ml at 0.3 g supplementation of sunflower, soybean, and mustard oil per litre of the media, respectively at 18 h incubation period. The present study indicates the Butyrivibrio fibrisolvens bacteria involves in the biohydrogenation process, and LA rich sunflower meal can be used to improve the CLA production in rumen and thereby increasing the CLA concentration of milk.

Keywords: Butyrivibrio fibrisolvens, CLA, fatty acids, sunflower oil

Procedia PDF Downloads 349

Authors: Nalini Murugaiyah

Abstract:

Indigenous students require a positive school environment where meaningful learning ought to be there to minimise myriad challenges. Therefore, Orang Asli student’s school environment should be culturally responsive and equipped with student-centred activities or provide constructively designed curriculum and pedagogy. This study sought to extend the knowledge of culturally responsive school leadership practises which relevant and responsive to Orang Asli students through th lens of a theoretical framework, Culturally Responsive School Leadership. The aim of the proposed study is to examine and understand the real-world application of leadership practices that are relevant and responsive to Orang Asli students in Malaysia. This study will also include the often-voiceless voices’ of Orang Asli students, parents, and community leaders to gain a deeper understanding of the process and experience of engaging in culturally responsive school leadership. The study will explore the differences between school leaders, teachers, parents, and community leaders in relation to culturally responsive school environment, non-Orang Asli school leaders’ and teachers’ support to the needs of Orang Asli children, children’s perspectives of teachers’ practices in the classroom align with their culture; and, the demonstration of teacher’s culturally responsive behaviour in the classroom. A basic qualitative study is the proposed research design for this study, and the data is collected through semi-structured interviews and focus group interviews. This qualitative research is designed to gain in-depth knowledge about how the principal’s leadership is culturally responsive towards the school environment, which will improve the quality of education received by the Orang Asli community in Malaysia, hence reducing the drop-out rates in Orang Asli students.

Keywords: indigenous leadership, equity, inclusion, policy

Procedia PDF Downloads 35

Authors: Hamid Mustafa, Heather J. Huson, Adeela Ajmal, Kim Euisoo, Tad S. Sonstegard

Abstract:

In this study, 67 animals, representing six different cattle breeds of Pakistan, were genotyped with the Bovine high density (777K) SNP Beadchip. These include 13 Sahiwal, 09 Red Sindhi, 13 Tharparkar, 08 Achi, 13 Cholistani and 10 Dhanni cattle breeds. Analysis of 500, 939 SNP markers revealed that the mean minor allele frequency (MAF) was 0.21, 0.22, 0.18, 0.23, 0.22 and 0.22 for Sahiwal, Red Sindhi, Tharparkar, Achi, Cholistani and Dhanni respectively. Significant differences of minor allele frequency (MAF) were observed between the indigenous Pakistani cattle population (P<0.001). Across these Pakistani cattle breeds, a common variant MAF (≥0.10 and ≤0.5) accounted for an overall estimated 75.71 % of the 500,939 SNPs and on the average 19.58 % of the markers were monomorphic. Mean observed (HO) and expected (HE) heterozygosities were 0.656 and 0.638, respectively. This primarily study of Pakistani indigenous cattle breeds indicate that this level of SNPs variation can potentially be used for genomic studies for future breeding plans and for farm animal conservation strategies.

Keywords: Pakistan, cattle, minor allele frequency, SNP, variation

Procedia PDF Downloads 617

Authors: M. Lizzy Madiwani, Ignatious Ncube, Evelyn Madoroba

Abstract:

This study was designed to determine the distribution of pathogenic bacteria in household raised chickens and study their virulence and antibiotic profiles. For this purpose, 40 chickens were purchased from families in the Capricorn district and sacrificed for sampling. Tissues were cultured on different bacteriological media followed by biotyping using Matrix-assisted Laser Desorption Ionization-time of Flight (MALDI-TOF). Disk diffusion test was performed to determine the antibiotic susceptibility profiles of these bacteria. Out of a total of 160 tissue samples evaluated, E. coli and Salmonella were detected in these tissues. Furthermore, determination of the pathogenic E. coli and Salmonella strains at species level using primer sets that target selected genes of interest in the polymerase chain reaction (PCR) assay was employed. The invA gene, a confirmatory gene of Salmonella was detected in all the Salmonella isolates. The study revealed that there is a high distribution of Salmonella and pathogenic E. coli in these chickens. Therefore, further studies on identification at the species level are highly recommended to provide management and sanitation practices to lower this prevalence. The antimicrobial susceptibly data generated from this study can be a valuable reference to veterinarians for treating bacterial diseases in poultry.

Keywords: antimicrobial, Escherichia coli, pathogens, Salmonella

Procedia PDF Downloads 96

Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

Abstract:

Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed, with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

Procedia PDF Downloads 587

Authors: A. M. Isakhanyan, F. N. Tkhruni, N. N. Yakimovich, Z. I. Kuvaeva, T. V. Khachatryan

Abstract:

Introduction: At present, the simbiotics based on different genera and species of lactic acid bacteria (LAB) and yeasts are used. One of the basic properties of probiotics is presence of antimicrobial activity and therefore selection of LAB and yeast strains for their co-cultivation with the aim of increasing of the activity is topical. Since probiotic yeast and bacteria have different mechanisms of action, natural synergies between species, higher viability and increasing of antimicrobial activity might be expected from mixing both types of probiotics. Endemic strains of LAB Enterococcus faecium БТK-64, Lactobaccilus plantarum БТK-66, Pediococcus pentosus БТK-28, Lactobacillus rhamnosus БТK-109 and Kluyveromyces lactis БТX-412, Saccharomycopsis sp. БТX- 151 strains of yeast, with probiotic properties and hight antimicrobial activity, were selected. Strains are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB and yeast strains were isolated from different dairy products from rural households of Armenia. The genotyping by 16S rRNA sequencing for LAB and 26S RNA sequencing for yeast were used. Combined cultivation of LAB and yeast strains was carried out in the nutrient media on the basis of milk whey, in anaerobic conditions (without shaker, in a thermostat at 37oC, 48 hours). The complex preparations were obtained by purification of cell free culture broth (CFC) broth by the combination of ion-exchange chromatography and gel filtration methods. The spot-on-lawn method was applied for determination of antimicrobial activity and expressed in arbitrary units (AU/ml). Results. The obtained data showed that at the combined growth of bacteria and yeasts, the cultivation conditions (medium composition, time of growth, genera of LAB and yeasts) affected the display of antimicrobial activity. Purification of CFC broth allowed obtaining partially purified antimicrobial complex preparation which contains metabiotics from both bacteria and yeast. The complex preparation inhibited the growth of pathogenic and conditionally pathogenic bacteria, isolated from various internal organs from diseased animals and poultry with greater efficiency than the preparations derived individually alone from yeast and LAB strains. Discussion. Thus, our data shown perspectives of creation of a new class of antimicrobial preparations on the basis of combined cultivation of endemic strains of LAB and yeast. Obtained results suggest the prospect of use of the partially purified complex preparations instead antibiotics in the agriculture and for food safety. Acknowledgments: This work was supported by the RA MES State Committee of Science and Belarus National Foundation for Basic Research in the frames of the joint Armenian - Belarusian joint research project 13РБ-064.

Keywords: co-cultivation, antimicrobial activity, biosafety, metabiotics, lactic acid bacteria, yeast

Procedia PDF Downloads 313

Authors: Ayodeji O. Falade, Leonard V. Mabinya, Uchechukwu U. Nwodo, Anthony I. Okoh

Abstract:

Given the high-utility of peroxidase, its production in large amount is of utmost importance. Over the years, actinomycetes have been the major peroxidase-producing bacteria. Consequently, other classes of bacteria with peroxidase production potentials are underexplored. This study, therefore, sought to enhance peroxidase production by a Raoultella species, a new ligninolytic proteobacteria strain, by determining the optimum culture conditions (initial pH, incubation temperature and agitation speed) for peroxidase production under submerged fermentation using the classical process of one variable at a time and supplementing the fermentation medium with some lignin model and inorganic nitrogen compounds. Subsequently, the time-course assay was carried out under optimized conditions. Then, some agricultural residues were valorized for peroxidase production under solid state fermentation. Peroxidase production was optimal at initial pH 5, incubation temperature of 35 °C and agitation speed of 150 rpm with guaiacol and ammonium chloride as the best inducer and nitrogen supplement respectively. Peroxidase production by the Raoultella species was optimal at 72 h with specific productivity of 16.48 ± 0.89 U mg⁻¹. A simultaneous production of a non-peroxide dependent extracellular enzyme which suggests probable laccase production was observed with specific productivity of 13.63 ± 0.45 U mg⁻¹ while sawdust gave the best peroxidase yield under solid state fermentation. In conclusion, peroxidase production by the Raoultella species was increased by 3.40-fold.

Keywords: enzyme production, ligninolytic bacteria, peroxidase, proteobacteria

Procedia PDF Downloads 250

Authors: Emiliano Castillo

Abstract:

The aim of this research is to analyze the origins, the development and possible outcomes of the environmental conflict between grassroots organizations, indigenous communities, Kinder Morgan Corporation, and the Canadian government over the Trans Mountain pipeline expansion in Burnaby, British Columbia, Canada. Building on the political ecology and the environmental justice theoretical framework, this research examines the impacts and risks of tar sands extraction, production, and transportation on climate change, public health, the environment, and indigenous people´s rights over their lands. This study is relevant to the environmental justice and political ecology literature because it discusses the unequal distribution of environmental costs and economic benefits of tar sands development; and focuses on the competing interests, needs, values, and claims of the actors involved in the conflict. Furthermore, it will shed light on the context, conditions, and processes that lead to the organization and mobilization of a grassroots movement- comprised of indigenous communities, citizens, scientists, and non-governmental organizations- that draw significant media attention by opposing the Trans Mountain pipeline expansion. Similarly, the research will explain the differences and dynamics within the grassroots movement. This research seeks to address the global context of the conflict by studying the links between the decline of conventional oil production, the rise of unconventional fossil fuels (e.g. tar sands), climate change, and the struggles of low-income, ethnic, and racial minorities over the territorial expansion of extractive industries. Data will be collected from legislative documents, policy and technical reports, scientific journals, newspapers articles, participant observation, and semi-structured interviews with representatives and members of the grassroots organizations, indigenous communities, and Burnaby citizens that oppose the Trans Mountain pipeline. These interviews will focus on their perceptions of the risks of the Trans Mountain pipeline expansion; the roots of the anti-tar sands movement; the differences and dynamics within the movement; and the strategies to defend the livelihoods of local communities and the environment against tar sands development. This research will contribute to the understanding of the underlying causes of the environmental conflict between the Canadian government, Kinder Morgan, and grassroots organizations over tar sands extraction, production, and transportation in Burnaby, British Columbia, Canada. Moreover, this work will elucidate the transformations of society-nature relationships brought by tar sands development. Research findings will provide scientific information about how the resistance movement in British Columbia can challenge the dominant narrative on tar sands, exert greater influence in environmental politics, and efficiently defend Indigenous people´s rights to lands. Furthermore, this research will shed light into how grassroots movements can contribute towards the building of more inclusive and sustainable societies.

Keywords: environmental conflict, environmental justice, extractive industry, indigenous communities, political ecology, tar sands

Procedia PDF Downloads 258

Authors: C. W. Kan, Y. L. Lam

Abstract:

Cotton textile has large specific surfaces with good adhesion and water-storage properties which provide conditions for the growth and settlement of biological organisms. In addition, the soil, dust and solutes from sweat can also be the sources of nutrients for microorganisms [236]. Generally speaking, algae can grow on textiles under very moist conditions, providing nutrients for fungi and bacteria growth. Fungi cause multiple problems to textiles including discolouration, coloured stains and fibre damage. Bacteria can damage fibre and cause unpleasant odours with a slick and slimy feel. In addition, microbes can disrupt the manufacturing processes such as textile dyeing, printing and finishing operations through the reduction of viscosity, fermentation and mold formation. Therefore, a large demand exists for the anti-microbially finished textiles capable of avoiding or limiting microbial fibre degradation or bio fouling, bacterial incidence, odour generation and spreading or transfer of pathogens. In this review, the main strategy for cotton textile will be reviewed. In the beginning, the classification of bacteria and germs which are commonly found with cotton textiles will be introduced. The chemistry of antimicrobial finishing will be discussed. In addition, the types of antimicrobial treatment will be summarized. Finally, the application and evaluation of antimicrobial treatment on cotton textile will be discussed.

Keywords: antimicrobial, cotton, textile, review

Procedia PDF Downloads 341

Authors: M. G. Shehata, S. A. El Sohaimy, Marwa M. Abu-Serie, Nourhan M. Abd El-Aziz

Abstract:

Probiotic strains can potentially be used as bio-preservatives and functional food supplement. Eight lactic acid bacteria strains (LAB) Lactobacillus brevis NRRL B-4527; Streptococcus thermophilus BLM 58; Pediococcusacidilactici ATCC 8042; Lactobacillus rhamnosus CCUG 1452; Lactobacillus curvatus ATCC 51436; Lactococcuslactis sub sp. lactisDSM 20481; Lactobacillus plantarum DMSZ 20079 and Lactobacillus plantarumTF103 were selected to screen the antioxidant, anticancer potential and probiotic properties. LAB strains exhibited good probiotic, antioxidant properties and showed antagonistic activity against food-borne pathogenic (Bacillus subtilis DB 100 host; Candida albicans ATCCMYA-2876; Clostridium botulinum ATCC 3584; Escherichia coli BA 12296; Klebsiellapneumoniae ATCC12296; Salmonella senftenberg ATCC 8400 and Staphylococcus aureus NCTC 10788). Further, in vitro probiotic properties of eight strains displayed excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, in vitro adhesion ability for HT-29 cell line. The antioxidant effect of intracellular and cell-free extract of lactic acid bacteria strains was evaluated by various antioxidant assays, namely, resistance to hydrogen peroxide, DPPH radical scavenging, ABTS radical scavenging, and hydroxyl radical scavenging (HRS). The results showed that intracellular and cell-free supernatant of S. Thermophilus BLM 58, L. lactissubsp.lactis DSM 20481, P. acidilactici ATCC 8042, L. brevis NRRL B-4527 strains possess excellent antioxidant capacity. The intracellular of S. Thermophilus BLM 58 and P. acidilactici ATCC 8042 also showed excellent anticancer activity against Caco-2, MCF-7, HepG-2, and PC-3. Antioxidative property of selected lactic acid bacteria strains would be useful in the functional food manufacturing industry. They could beneficially affect the consumer by providing dietary source of antioxidants.

Keywords: anticancer activity, antioxidant activity, functional food, lactic acid bacteria, probiotic

Procedia PDF Downloads 201

Authors: Karl D. Humm

Abstract:

The United Nations’ Community-based REDD+ (Reducing Emissions from Deforestation and forest Degradation) (CBR+) is a programme that directly finances grassroots climate change mitigation strategies that uplift Indigenous Peoples (IPs) and other marginalised groups. A pilot for it in six countries was developed in response to criticism of the REDD+ programme for excluding IPs from dialogues about climate change mitigation strategies affecting their lands and livelihoods. Despite the pilot’s conclusion in 2017, no complete report has yet been produced on the results of CBR+. To fill this gap, this study investigated the experiences with involving IPs in the CBR+ programmes and local projects across all six pilot countries. A literature review of official UN reports and academic articles identified challenges and successes with IP participation in REDD+ which became the basis for a framework guiding data collection. A mixed methods approach was used to collect and analyse qualitative and quantitative data from CBR+ documents and written interviews with CBR+ National Coordinators in each country for a cross-country comparative analysis. The study found that the most frequent challenges were lack of organisational capacity, illegal forest activities, and historically-based contentious relationships in IP and forest-dependent communities. Successful programmes included IPs and incorporated respect and recognition of IPs as major stakeholders in managing sustainable forests. Findings are summarized and shared with a set of recommendations for improvement of future projects.

Keywords: climate change, forests, indigenous peoples, REDD+

Procedia PDF Downloads 101

Authors: Disaya Jaroensattayatham

Abstract:

Nowadays, infectious diseases are prevalent and severe health problems as they render the increment of casualty, illness, and global economic recession. Along with the emergence of antimicrobial resistance, the potency of typically used antibiotics can be affected to a considerable degree. As a result, unorthodox antibiotics have become an urgent issue in the pharmaceutical field. Piper betle L., known as betle leaf, has been used for many purposes, such as a traditional home remedy, and has shown its ability in inhibiting bacteria as well as fungus. Thus, in this study, the investigation of antimicrobial activities of the Piper betle L. extracts was carried out using the Agar disk-diffusion method and Broth microdilution, aiming to evaluate and determine its efficacy to inhibit bacterial and fungal growth of Staphylococcus aureus, Salmonella typhi, and Candida albicans. In the agar disk-diffusion test, the extracts of Piper betle L. gave the maximum zone of inhibition of 15.1 mm (S. aureus), 7.7 mm (S. typhi), and 11.7 mm (C. albicans), while its MIC values were 1000 µg/ml in S. aureus and greater than 2000 µg/ml in S. typhi and C. albicans. According to the results, the Piper betle L. obtains an antimicrobial activity and shows a higher effect towards gram-positive bacteria than gram-negative bacteria. To determine the mechanism behind its ability, more research is needed to be performed in the future.

Keywords: antimicrobial activity, Candida albicans, Piper betle L., Salmonella typhi, Staphylococcus aureus

Procedia PDF Downloads 146