Browsing by Author "Bethel Sitotaw"

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    Screening of Endophytic Bacteria Isolated from Selected Plants in Hawassa for Production and Characterization Amylase
    (Addis Ababa University, 2024-06) Bethel Sitotaw; Asnake Desalegn; Fitsum Tigu
    Amylases are crucial enzymes in the global industry, constituting 25% of enzyme production. Endophytes, known for producing bioactive compounds and enzymes, offer promise for industrial applications. The aim of the study was to screen amylase producing endophytic bacteria, to produce amylase under controlled laboratory conditions and characterize the enzyme. Purposive sampling was used to select the study areas as well as the plants screened for amylase producing endophytic bacteria. After collection the plant samples were washed and surface sterilized before isolation of the endophytic bacteria. The endophytic bacteria were isolated then screened for amylase production on starch agar media and purified. The pure isolates were characterized using morphological and biochemical test. Amylase was produced through submerged fermentation, and its production was optimized using different carbon and nitrogen sources, pH and temperatures. Ammonium sulphate precipitation and dialysis were used for partial purification and the partially purified enzyme was assayed for its activity using the dinitrosalicylic acid (DNS) method. Out of 60 endophytic bacteria isolated from the plants, 32 demonstrated amylase production. From these, 16 pure isolates were found to be the most efficient amylase producers and 7 best amylase producing isolates based on submerged fermentation. Among these top 16 producers, 12 were Gram-positive bacteria and 4 were Gramnegative bacteria.The 7 isolates were identified to species level by rRNA gene sequence analysis. Then 3 isolates were selected for further purification and characterization. Diameter (Mean ± SD) of clear zone on starch agar ranged from 4.48 ± 0.54 for isolate I10L2 from Enset to 11.53 ± 0.34 for isolate D10F2 from Datura. The highest (p < 0.05) amylase production was recorded for isolate D10L2 when glucose or starch was used as carbon source. The highest amylase production was recorded for all isolates in the presence of ammonium sulfate and the lowest in the presence of tryptone as nitrogen source. The optimum temperature for production of amylase was 30 OC, but differences in the optimum pH ranging from pH 5 to pH 8 were recorded based on the isolates preferences. After partial purification the highest enzyme activity was recorded at pH 7 and pH 9, 50 OC, and 0.5% and 1% calcium ion (Ca 2+) concentrations. The maximum activity in the presence of 1mM Ca2+ was about 16.5 U/mL. These findings highlight the potential of endophytic bacteria for amylase production. With further optimization and scale up, the amylase can be produced in large quantities and for specific applications.
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    Screening of Endophytic Bacteria Isolated from Selected Plants in Hawassa for the Production and Characterization of Amylase
    (Addis Ababa University, 2024-06) Bethel Sitotaw; Asnake Desalegn; Fitsum Tigu (PhD)
    Amylases are crucial enzymes in the global industry, constituting 25% of enzyme production. Endophytes, known for producing bioactive compounds and enzymes, offer promise for industrial applications. The aim of the study was to screen amylase producing endophytic bacteria, to produce amylase under controlled laboratory conditions and characterize the enzyme. Purposive sampling was used to select the study areas as well as the plants screened for amylase producing endophytic bacteria. After collection the plant samples were washed and surface sterilized before isolation of the endophytic bacteria. The endophytic bacteria were isolated then screened for amylase production on starch agar media and purified. The pure isolates were characterized using morphological and biochemical test. Amylase was produced through submerged fermentation, and its production was optimized using different carbon and nitrogen sources, pH and temperatures. Ammonium sulphate precipitation and dialysis were used for partial purification and the partially purified enzyme was assayed for its activity using the dinitrosalicylic acid (DNS) method. Out of 60 endophytic bacteria isolated from the plants, 32 demonstrated amylase production. From these, 16 pure isolates were found to be the most efficient amylase producers and 7 best amylase producing isolates based on submerged fermentation. Among these top 16 producers, 12 were Gram-positive bacteria and 4 were Gramnegative bacteria.The 7 isolates were identified to species level by rRNA gene sequence analysis. Then 3 isolates were selected for further purification and characterization. Diameter (Mean ± SD) of clear zone on starch agar ranged from 4.48 ± 0.54 for isolate I10L2 from Enset to 11.53 ± 0.34 for isolate D10F2 from Datura. The highest (p < 0.05) amylase production was recorded for isolate D10L2 when glucose or starch was used as carbon source. The highest amylase production was recorded for all isolates in the presence of ammonium sulfate and the lowest in the presence of tryptone as nitrogen source. The optimum temperature for production of amylase was 30 OC, but differences in the optimum pH ranging from pH 5 to pH 8 were recorded based on the isolates preferences. After partial purification the highest enzyme activity was recorded at pH 7 and pH 9, 50 OC, and 0.5% and 1% calcium ion (Ca 2+) concentrations. The maximum activity in the presence of 1mM Ca2+ was about 16.5 U/mL. These findings highlight the potential of endophytic bacteria for amylase production. With further optimization and scale up, the amylase can be produced in large quantities and for specific applications.
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    The Effect of Community Engagement, Capacity-Building, and Monitoring and Evaluation on The Sustainability of Communitybased Projects: A Case Study of Family Guidance Association of Ethiopia South Area Office (Fgae-Sao)
    (Addis Ababa University, 2024-10) Bethel Sitotaw; Fesseha Afewerk (PhD)
    Community-based projects (CBPs) are a type of community-based project that has a significantimpact on society. The sustainability of CBPs is influenced by a variety of factors such as community engagement, capacity building, and monitoring and evaluation (M&E) practices. Byusing the Family Guidance Association of Ethiopia South Area Office as the case study organization, the study aimed to analyze factors that impact the sustainability of communitybasedprojects. The study aimed to determine the relationship between community engagement, capacity building, monitoring, and evaluation of community-based project sustainability. Theresearch design used in this study was descriptive with a mixed-method approach. Quantitative data was gathered through the administration of close-ended Likert scale questionnairescontaining additional open-ended questions while qualitative data was gathered through interviews. The target population was the administrative staff, program staff, and members of thecommunity in the organization. The Statistical Package for Social Sciences (SPSS) analyzed thequantitative data and displayed the results in tables using frequencies, percentages, means, andstandard deviations. Pearson’s correlation was used to determine the relationship between thevariables. Interview data were analyzed using thematic analysis. The results showed thatCommunity engagement is perceived at a moderate level, highlighting the need for enhanced communication and inclusive decision-making. A robust positive correlation between communityparticipation and project sustainability is supported by regression analysis, indicating a significant positive impact.Capacity-building activities also exhibit a moderate presence and positively correlate with project sustainability. Regression analysis reveals that capacity building significantly influencesproject outcomes, emphasizing its role despite some relationships not achieving statistical significance.Systematically implemented Monitoring and evaluation practices demonstrate a strong positivecorrelation with project sustainability. Regression analysis underscores M&E's critical role as apredictor of positive project outcomes. FGAE employs diverse funding strategies and incomegeneratingactivities for sustainability, yet varying confidence levels suggest a need for moreconsistent long-term planning. These findings contribute to existing knowledge on achievinglong-term sustainability in community-based projects. Keywords: community-based projects, community involvement, capacity building, monitoringand evaluation, project sustainability, sustainable development