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Characterization of Antioxidant Agent Producing Bacteria Form Intestinal Flora F Farm Animals

Thesis Info

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Author

Noureen, Saleha

Program

PhD

Institute

University of the Punjab

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Microbiology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10775/1/Saleha_Noureen_Microbiology_2019_UoP_Punjab_19.03.2019.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725679157

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Excessive reactive oxygen species (ROS) damage cell membrane, cellular components and cause oxidative damage. They also lead to various human disorders such as diabetes, cancer, Alzheimer, rheumatoid arthritis and inflammation. The body possesses antioxidative defense and repair mechanisms to protect against such damages. But sometime this system is unable to prevent entire damage caused by ROS. In such conditions, supplementation of exogenous antioxidant has considered better options. Several synthetic antioxidants (butylated hydroxy-toluene (BHT), propyl-gallate and butylated hydroxyl-anisole (BHA) are extensively used, but their safety is questioned due to recent reports on their toxicity. In this scenario, exploration of natural and safer antioxidant from bio-resources and their replacement with synthetic antioxidant is highly commended. Lactic acid bacteria (LAB) are an emerging natural source of cellular antioxidants. LAB are diversely dispersed in nature. Previously LAB isolate from dairy products and some from human fecal matter. Information about few isolated strains of LAB is available from farm animals. The aims of this work were (a) to explore lactic acid bacteria from farm animals for their antioxidant potential. b) to check probiotic potential of strains with promises antioxidant capability and identify potential competent strain (c) to investigate in vivo antioxidant ability of competent strain (d) to perform its safety assessment and finally e) the partially purify secondary metabolite responsible for the antioxidant property of the strain. The strains were identified using conventional microbiological and molecular methods. The antioxidant scavenging assay against three radicals (DPPH, superoxide anion and OH radical) and inhibition of lipid peroxidation were used to evaluate the antioxidant potential in vitro. Twenty three strains were investigated in intact cells, the supernatant and cell lysate. Highest antioxidant activity was noticed in intact cell and supernatant while weak activity was observed in cell lysate. The promiser antioxidant activity in four assays was recorded. Seven out of twenty three LAB strains displayed above 60% antioxidant activity in all four tests and were selected for further study. The probiotic potential of the selected seven strains was assessed through tolerance against different pH and temp, resistance to bile salt, sodium chloride salt and simulated gastric intestinal environment. The strains were able to tolerate temperature (30-55C), pH (2-10), bile salt (up to 2%), sodium chloride (up to 8%) and simulated gastrointestinal environment (≥ 80% survival) proving their probiotic nature. In addition the strains were noticed to pose least threat to eukaryotic cells identified by very weak hemolytic activity (0.2-1.2%) against host erythrocytes. The seven strains were identified as Enterococcus rati (An8), Lactobacillus brevis (An13, An15 An26, An27 and An28) and Pediococcus acidilactici (An17) on the basis of 16S rRNA gene sequencing. They were awarded accession no are MF183967, MH185807, MH185808, MG882401, MG882402 and MG000874 from NCBI Gene bank. These strains are also deposited in Fungal Culture Bank for the use of other researchers. However, among all the performance of An28 was excellent. Therefore, this strain was chosen for in vivo validation of its antioxidant potential. The strain An28 was identified as L. brevis MG000874. The Efficacy of this strain was checked by analyzing selected oxidative stress parameters (SOD, CAT, GST and GSH) and general health in d-galactose model of oxidative stress. Two experiments were performed in this regard. A pilot study was conducted to determine the dose of d-galactose required to induce oxidative stress in mice. Dose of 150mg/Kg BW/day for sixty days could successfully induced oxidative stress. The oxidative stress was identified by depletion in SOD, CAT and GSH and elevation of GST elevation for eight weeks. In the main trial, the prophylactic effect of L. brevis MG000874 supplementation worked out. Physical sign and symptoms as well as estimation of oxidative stress indicators of liver, kidney and serum were measured. The analysis of SOD, CAT, GST and GSH provided evidence that L. brevis MG000874 can resist the occurrence of oxidative stress. The continue feeding of L. brevis MG000874 @109cfu/ml for a period of 8 weeks did not adversely affect the general health of animals.This strain was fed to the Mus musculus for eight weeks. After eight weeks of treatment, liver and kidney homogenate were used for analysis of known markers of toxicity, including ALT, AST, ALP, T. bilirubin, Albumin and protein from liver and Urea and Creatinine test from kidney. In addition histological studies on liver and kidney were also carried out on the size of central vein, sinusoids and hepatocytes of liver and glomerulus, corpuscles, PCT and DCT in kidney were taken into account. LFT and RFT analysis revealed the safe nature of L. brevis MG000874 which was also supported by histological findings. In the end the metabolites released in the cell free supernatant by L. brevis MG000874 were partially purified. In this procedure, the metabolites having antioxidant property were separated through thin layer chromatography (TLC). These separated antioxidant components were analyzed in High performance liquid chromatography (HPLC) and Attenuated Total Reflectance Infrared (ATR-IR) for determination of purity and the chemical nature of pure compound. The analysis revealed that the component was aldehyde in nature.It is concluded that L. brevis MG000874 produce antioxidative compounds of aldehyde nature that may serve as a novel agent for controlling the oxidative stress in host. Live L. brevis MG000874 or its metabolites may be included in the list of potential probiotics and prebiotics respectively.
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شبِ برات اور آتشبازی کی قبیح رسم

شب برات اور آتش بازی کی قبیح رسم
انسان کی ہمیشہ سے خواہش رہی ہے کہ کام تھوڑ ا کر نا پڑے اور اس کی مزدوری اور اُجرت زیادہ مل جائے۔ اللہ تعالیٰ نے بھی کچھ اوقات ایسے مقرر کیے ہیں کہ اس میں دین اسلام پر کار بند شخص تھوڑی سی عبادت کر کے ڈھیروں ثواب کما سکتے ہیں۔ ان اوقات میں لیلۃ القدر یعنی شب برات بھی ہے جو پندرہ شعبان المعظم کی رات ہے اور کروڑوں مسلمان اس رات میں شبِ بیدار ی کر کے اپنے پروردگار کے سامنے سر بسجدہ ہوتے ہیں اور اپنے گناہوں کی معافی کے طلبگار ہوتے ہیں۔ چنانچہ قرآن مجید میں ہے کہ:۔
’’حم قسم ہے کتاب مبین کی بیشک ہم نے اتارا ہے اسے ایک برکت والی رات میں بے شک ہم ڈر سنانے والے ہیں، اس رات ہرحکمت والا کا م بانٹ دیا جا تا ہے۔‘‘
تشریح! یہاں کتاب مبین سے مراد کلام اللہ یعنی قرآنِ مجید فرقانِ حمید ہے اگر چہ بعض مقامات پر اس سے مرادلوح محفوظ بھی ہے۔ نزولِ قرآن کی رات کی عظمت و فضیلت کو ظاہر کرنے لیے اللہ تعالیٰ نے قرآنِ مجید کی قسم ارشاد فرمائی۔ لیلۃ القدر برکتوں والی رات اس رات کے تعین میں مختلف اقوال ہیں لیکن عام طور پرد وقول زیادہ مشہور ہیں ایک تو یہ کہ اس سے مراد لیلۃ القدر ہے جو ماہ ِرمضان شریف میں آتی ہے۔ دوسرا قول یہ ہے کہ اس سے مراد شب ِبرات ہے جو شعبان المعظم کی پندرھویں رات ہے جیسا کہ تفسیر مظہری میں ہے۔ حضرت عکرمہصنے کہا کہ یہ پندرھویں شعبان کی رات ہے جس میں سال بھر کے امور لکھ دیئے جاتے ہیں جنہیں مرنا ہوتا ہے انہیں زندوں کی فہرست سے نکال دیا جاتا ہے پھر ان میں نہ زیادتی کی جاتی ہے نہ کمی۔

The Prophet Muḥammad (S. A. W) : A Universal Messenger of Peace (An Overview of the Prophet Muḥammad (SAW) ’s Relations With the Non-Muslims

Islām, as the religion of peace and mercy, teaches and preaches peace, justice, tolerance, goodwill and equality among the human beings. Being the trustee of Allāh, man has been prescribed some requirements for the regulation of human conduct. The Prophet Muḥammad (r) is the personification of these teachings and commandments of Allāh. His conduct is “the Perfect Example” and “the Role Model” to be followed by human beings. He has been titled in The Qur’ān as (al-Raḥmah) the mercy for all the worlds. Whatever aspect of the Prophet Muḥammad (r) ’s life is investigated, it is proved to be unbiased and stands as a hallmark for the universal peace, justice, tolerance and equality, not only for the believers, but, for the non-Muslims, even for the worst enemies, as well. The first part of this paper describes the concept of peace in Islām and the role of the Prophet Muḥammad (r) as the messenger of the religion of peace. The second part of the paper focuses on the characteristics of the Prophet Muḥammad (r) as the messenger of peace in every aspect of life. While the third part of the paper is about the study of the historical facts, which discuss the life of the Prophet Muḥammad (r) as a historical figure and there is no doubt in his personality and character.

Proactive Strategies to Improve Link Stability under Topology Changes in Mobile Adhoc Networks

Improving link stability under topological changes arise due to link/node failures under uniform speeds and random trajectories of mobile nodes, is a significant problem that must be addressed to improve routing performance in mobile adhoc networks (MANETs). Under random characteristics of wireless channels, there must be a unified model based on proactive strategies to improve link stability under topological changes for stable and robust communications among neighboring nodes. This thesis addresses the problem of link stability under topological changes by presenting proactive strategies for faster failure detection, robustly maintaining link stability, effective link stability estimation, and efficient traffic estimation. First, a novel faster failure detection (FFD) strategy is proposed for link re-connectivity under link/node failures using a Hello messaging scheme. FFD is modeled through a Continuous Time Markov Chain (CTMC) based approach to proactively detect link/node failures under topological changes in MANETs. The FFD strategy minimizes delay incurred as a result of link re-connectivity, optimizes adjacency establishments among neighboring nodes, and enables energy efficiency to improve network connectivity. Second, a novel dynamic link connectivity (DLC) strategy for maintaining link stability among neighboring nodes. DLC is modeled through CTMC by taking into account continuous time analysis of dynamic link connectivity and stationary probability distribution between neighboring node pairs. The DLC strategy examines stability of network nodes and calculates probabilities of connection and disconnection states on the basis of transition rate estimation between these states. The DLC strategy robustly maintains link connectivity for future communications among neighboring nodes inside a cluster. Third, novel link connectivity metrics (LCM) and path distribution analysis (PDA) strategies are presented for effective link stability estimation in MANETs. LCM and PDA are modeled using CTMC under random movement of network nodes and topology trigxxii gered path duration statistics as a function of effective link stability estimation. The LCM and PDA strategies collectively enhance robustness and stability of a network by making it less prone to connectivity failures for future communications among network nodes. Finally, the fourth contribution is a novel efficient traffic estimation (ETE) strategy using Hello messaging among neighboring nodes inside a cluster. ETE is modeled through periodic exchange of transmitted Hello messages under local link connectivity between neighboring node pairs, which follows a CTMC model. The ETE strategy estimates optimum Hello messages traffic through local link connectivity. Simulation and analytical results demonstrate that the proposed strategies improve link stability under topological changes and can be deployed as effective tools for maximizing routing performance in MANETs.