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Home > Molecular Detection and Charaterization of Escherichia Coli Isolated from Raw Cow Milk Samples Collected from District Muzaffargarh

Molecular Detection and Charaterization of Escherichia Coli Isolated from Raw Cow Milk Samples Collected from District Muzaffargarh

Thesis Info

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External Link

Author

Saadia Niaz

Institute

Virtual University of Pakistan

Institute Type

Public

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Software Engineering

Language

English

Link

http://vspace.vu.edu.pk/detail.aspx?id=293

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676721015849

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Milk is generally considered as highly nutritious and useful food for all age groups as well as inexpensive and widely available. However, its quality and microbial content are the features which have to be observed. The quality of milk depends on its biochemical composition and hygienic conditions during the collection of milk and its distribution. Milk itself acts as best medium for enhancing the growth and development of different types of microbes because of its composition and presence of water in large quantity. Milk sold in Muzaffargarh district was suspected to be contaminated because of repeated outbreaks of gastroenteritis. In this context, the research was designed to rule out molecular detection and quantitative analysis of Escherichia coli (E. coli) in the milk samples obtained from various tehsils of Muzaffargarh district. E. coli were considered as the most prevalent bacteria of milk and could be the most probable microbe causing this disease. In this research, 100 raw milk samples were gathered in a way that 10 samples each were collected from Jutoi, Alipur, Kotadu, Murad Abad, Choak Qureshi, Ahmadpur, Rohilan Wali, Shahjamal, Khanpur and Muzaffargarh city. Then the samples of milk were cultured on different culture media for bacterial segregation. Identification of bacterial specie was done through gram?s staining and properties of bacterial culture on different selective media. Biochemical tests were also performed which include catalase and coagulase tests. Final identification was performed through PCR and resolution of PCR products by gel electrophoresis. Antibiotic sensitivity test was also performed so as to confirm the susceptibility of E. coli regarding multiple antibiotics. Out of 10 samples collected each from Jutoi, Alipur, Kotadu, Murad Abad, Choak Qureshi, Ahmadpur, Rohilan Wali, Shahjamal, Khanpur and Muzaffargarh city, results had shown the presence of E.coli in 4(40%), 6(60%), 3(30%), 2(20%), 4(40%), 5(50%), 3(30%), 4(40%), 6(60%) and 5(50%) samples respectively. E. coli isolates were amplified by PCR based on 16S rRNA gene. Results of antibiotic sensitivity test revealed that E. coli isolates had shown resistance to amoxicillin (85%) and erythromycin (72%). However, they were found to be affected by azithromycin (53%), ciprofloxacin (86%), gentamicin (86%), norfloxacin (80%) and streptomycin (66%). Resistant pattern in relation to broad spectrum antibiotic (i.e., amoxycillin) points to a situation which should be considered carefully and suggests that indiscriminate use of antibiotics for precautionary or therapeutic purposes should be avoided as it could be the cause of increasing antimicrobial resistance.
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شفیق الرحمن قدوائی

شفیق الرحمن قدوائی
شفیق الرحمن مرحوم اگر چہ شہرت و ناموری کے عام معیار سے کو ئی بڑے آدمی نہ تھے مگر اپنے ایثار و قربانی، اخلاق وکردار، اخلاص و عمل اور خاموش اور بے لوث خدمات کے لحاظ سے بہت سے بڑے بڑے لیڈروں پر فائق تھے، جامعہ ملیہ کے لئے تو انھوں نے اپنی زندگی وقف کردی تھی اور سرد و گرم دور میں بھی اس سے جدا نہ ہو ئے، اور یہ کہنا غلط نہ ہوگا کہ جامعہ انہی کی محنت و جانفشانی کی بدولت زندہ رہ گیا، ظاہر وباطن دونوں میں مسلمان اور اپنے اوصاف کی بنا پر ہر جماعت میں مقبول تھے، کا نگریس اور حکومت دونوں کے سنجیدہ طبقہ میں ان کا بڑا وقار و وزن اور اخلاقی اثر تھا، مگر وہ اتنے بے لوث تھے کہ کبھی اس اثر سے فائدہ اٹھانے کی کوشش نہیں کی، ان کو بنیادی تعلیم کا عملی تجربہ تھا، اس کے وہ ماہر تھے، اس لئے یو این او کی جانب سے اس کام کے لئے انڈونیشیا بھیجے گئے تھے، ابھی وہ وہیں تھے کہ گذشتہ الیکشن میں کانگریس نے ان کو دہلی اسمبلی کے لئے مقرر ہو ئے، مگر اس سے بھی ان کا فائدہ اٹھانے کا موقع نہ مل سکا، تھوڑے ہی دنوں کے بعد بیمار پڑگئے، اور چند مہینے بیمار رہ کر ۳؍ اپریل کو انتقال کیا، انتقال کے وقت کل ۵۳ سال کی عمر تھی جو سیاست کی دنیا میں عین شباب کی عمرہے، مسلمانوں میں اب ایسے مخلص اور باعمل آدمی مشکل سے پیدا ہوں گے، اﷲ تعالیٰ اس پیکر اخلاص کو اپنی رحمت و مغفرت سے سرفراز فرمائے۔ (شاہ معین الدین ندوی، اپریل ۱۹۵۳ء)

 

مسالک کے مشترکات کے ادراک کی اہمیت: پاکستان کے تناظر میں تجزیاتی مطالعہ

Signaficance of the Understanding of Intra-faith Similerties: Analytical Study in the Context of Pakistan Muslims are commanded to foster unity as breaking into sects is forbidden by Allah. Islam teaches about broadness of vision and the emergence of different denominations in Islam is because of this broadness. There are different school of thoughts that emerged due to the broader perspective of Shar’ῑah rulings like Hanfῑ, Shᾱfῑ, Mᾱlikῑ and Hanblῑ, J'afrῑ etc despite that there is an essential unity in beliefs and practices among the Muslims. They all worship Allah, follow the last Messenger, Muhammad (ﷺ) and the last revelation Qur’an. They face the same Qibla while praying, prostrate to Allah five times a day, and believe in finality of prophet hood. Qur'an and Ahᾱdῑth are a source of jurisprudence for all Muslims. The difference between Muslims is in understanding and interpreting the Scripture and Ᾱhᾱdῑth of Prophet Muhammad (ﷺ) in the matters related with implementation of certain religious, social, political, and other duties. Islam rejects sectarianism, intolerance and extremism. Keeping in mind all of the above-mentioned points, in the article an attempt has been made to analyze the major challenges facing the intra-faith unity in Pakistan. The first is ignorance. Second is the role of media and scholars. Third is curriculum and fourth is intolerance. In the beginning the introduction of different schools of thought is given, and then forbearance demonstrated from the life of ‘Salaf Sᾱlihῑn’ has been described to establish an atmosphere of harmony in the present time, especially in Pakistan. The importance and significance of foundations of harmony is explained in such a manner that every Muslim should understand that the differences between the Muslims are very small, as they are only minor disagreements. Other than that, they are united in beliefs and practices. Finally, in the end, recommendations have been proposed.

Effect of Site-Directed Mutagenesis on the Efficiency of the Amylase Gene of B. Licheniformis

Bacillus α-amylases are widely used in industrial processes. Nevertheless, there is a need to improve the temperature tolerance of α-amylases as high temperature conditions are used in many industrial processes. In this study, various isolates of Bacillus licheniformis (B. licheniformis) were recovered from soil using potatoes (Solanum tumberosum) and sweet potatoes (Ipoema batatas) for baiting. Isolates were identified using common morphological techniques and biochemical tests with an API kit. B. licheniformis isolates for further study were selected on the basis of high α-amylase production. The α-amylase gene was cloned in plasmid pTBSG1 and characterized. Mutations to be introduced for enhancing α-amylase stability were predicted by in-silico design using various computational tools: Rosetta, the disulfide design server and the RosettaVIP tool. The disulfide design server identified five double mutants for mutagenesis to cysteine residues as follows: L64C/K80C, E119C/S130C, G433C/G474C, T139C/W157C and Y396C/D416C. A sixth mutant, V283I, was predicted with the RosettaVIP tool. The various mutants were prepared with the Phusion site-directed mutagenesis kit with the help of modified primers. Wild-type and mutants were then recombinantly expressed in E. coli BL21 (DE3) and characterized and compared to wild-type enzyme. Wild-type and mutant enzymes were pre-incubated at 45 ºC to 85 ºC and residual activities were found to not vary significantly till 55 ºC (P-value > 0.05). On the other hand, at 65 ºC, 75 ºC and 85 ºC, mutant V283I showed significantly improved thermostability (P-value < 0.05). Among the double mutants, G433C/G474C was found to have highest thermostability in terms of thermal inactivation (P < 0.05). This study shows that the rational design and introduction of substitution mutations leading to the formation of disulfide bonds and increased hydrophobic interactions has high potential for improving the thermal stability of α-amylases.