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Home > Genetic Diversity and Molecular Basis of Multidrug Resistance in Acinetobacter Baumannii Clinical Isolates

Genetic Diversity and Molecular Basis of Multidrug Resistance in Acinetobacter Baumannii Clinical Isolates

Thesis Info

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Author

Khurshid, Mohsin

Program

PhD

Institute

Government College University

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Microbiology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9832/1/Mohsin%20Khurshid_Microbio_GCU%28F%29_Main%20part.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726291680

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Acinetobacter baumannii is an increasingly important hospital-acquired Gramnegative bacterium that can thrive in the environment of healthcare facilities, and possess a significant public health concern. These features accompanied by its inherent capacity of resistance to antimicrobial agents, acquisition of diverse resistance mechanisms and the aptitude for epidemic spread greatly contribute to the success of A. baumannii as the most important nosocomial pathogen. The main aim of the present study was to investigate the molecular mechanisms underlying the multidrug-resistant phenotypes and the molecular epidemiology of this ignored pathogen of high clinical importance from Pakistan. A total of 319 A. baumannii isolates obtained from various clinical specimens were identified by routine microbiological procedures and further confirmed by multiplex PCR for the amplification of recA gene and internal transcribed spacer (ITS) region along with the amplification of blaOXA-51-like gene. The antimicrobial susceptibility pattern was determined through disc diffusion method and MIC was measured using agar dilution, broth microdilution and E-test® methods. The presence of genes encoding resistance to beta-lactams, 16S rRNA methylases, aminoglycoside modifying enzymes, fluoroquinolones, tetracyclines and sulfonamides were evaluated by PCR followed by sequencing. Repetitive extragenic palindromic PCR (REP-PCR) and multilocus sequence typing (MLST) was performed to investigate the genetic diversity. According to the results, the 96.6% isolates were multidrug-resistant (MDR) and 84.3% were extreme drug-resistant (XDR); 298 (93.4%) were resistant to carbapenems. The blaOXA-51was identified in all A. baumannii strains confirmed by multiplex PCR whereas the acquired blaOXA-23 gene was identified in 284 (89%) isolates. Higher rates of resistance were observed to the extended-spectrum cephalosporins including ceftazidime, cefotaxime, ceftriaxone and cefepime with MIC50 ≥ 128 μg/ml. The blaOXA-23 gene with an upstream ISAba1 was the foremost mechanism of carbapenem resistance that was found in 279 (87.5%) isolates and the blaNDM was found in only 7 strains belonging to a single MLST type. The genes encoding plasmid-mediated quinolone resistant were not detected in any isolate and the mutations in the gyrA and parC genes were the main underlying mechanism responsible for fluoroquinolone resistance. The 209 (65.5%) isolates were resistant to tetracyclines and 94.3% of these isolates were found positive for tetB gene. Among the sulfonamide resistance determinants, sul2 (85.2%) was the most common gene followed by sul1 (32.8%) whereas the combination of sul1 and sul2 genes was detected in 24.6% isolates. All the XIX isolates were found susceptible to polymyxins (polymyxin B and colistin) with MIC50 as 0.5 μg/ml as well as tigecycline with MIC50 (1 μg/ml). On the basis of REP-PCR the indigenous isolates were separated into 8 distant clones whereas MLST demonstrated the presence of seven already reported STs (ST642, ST589, ST2, ST600, ST338, ST103, and ST615) from different parts of world and eight new sequence type that were single or double locus variants to each other. The predominant STs i.e. ST642 and ST589 belonged to clonal complex I according to the Pasteur scheme and were found to harbor blaOXA-23 gene. The overall multidrug resistance was almost common among the isolates of already established STs whereas the isolates belonging to ST338 and the new STs were mainly susceptible. This dichotomy specifies the major selective advantage exerted by the antimicrobial resistance that drives the enduring clonal expansion of multidrug-resistant pathogens. The study revealed the alarming trends of multidrug resistance and substantial genetic diversity among A. baumannii clinical isolates from Pakistan. The differences in the distribution of various antimicrobial resistance mechanisms among various clones demonstrate the capacity of A. baumannii to acquire and express a wider range of resistance determinants. The deeper understanding of the genetic and biochemical basis of antibiotic resistance is of principal importance to design the policies for the effective control of emergence, spread, and development of innovative approaches for the therapeutic management of these multidrug-resistant pathogens.
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سید صدیق حسن

سید صدیق حسن
سخت افسوس ہے ۶؍ ستمبر کویوپی گورنمنٹ کے سب سے زیادہ سنیئر آئی سی ایس، ممبرریونیو بورڈ سیدصدیق حسن صاحب نے اچانک داعیِ اجل کولبیک کہا اور اس دنیا سے رخصت ہوگئے۔انتقال سے صرف پانچ روز پہلے یعنی یکم ستمبر کو عصر کی نماز کے بعد دارالعلوم ندوۃ العلماء لکھنؤ کی مجلسِ انتظامیہ کاجلسہ تھا، مرحوم سے وہاں ملاقات ہوئی،حسب عادت بڑے تپاک اور گرم جوشی سے ملے۔ جلسہ کے اختتام پر ہم سب کے ساتھ مسجد ندوہ میں مغرب کی نماز پڑھی، باہر نکلے توراقم الحروف اوردوسرے حضرات کے ساتھ دس پندرہ منٹ بات چیت کرتے رہے اور پھر مولانا عبدالماجدصاحب دریابادی کواپنے ساتھ لے کر کارمیں بیٹھ کر رخصت ہوگئے۔ اس وقت دیکھنے میں کافی تندرست اور ہشاش بشاش تھے اوراس بات کا وہم وگمان بھی نہیں ہوسکتا تھا کہ بس اب عالمِ آب وگِل میں پانچ دن کے مہمان ہیں۔ پاکستان میں ایک قریبی عزیز کاانتقال ہوگیاتھا، اُن کی تعزیت کرنے کی غرض سے اپنی بیوی اور بیٹی کے ساتھ لاہور جارہے تھے ،امرتسر پہنچ کر کسٹم وغیرہ کے مراحل سے گزرنے کے لیے ایک متعلق افسر کی میز کے سامنے جاکر کھڑے ہوئے اور جیب سے پاسپورٹ نکال کرافسر مذکور کی طرف بڑھارہے تھے کہ وقتِ موعود آپہنچا ،یک بیک دل کی حرکت بندہوگئی اور دھڑام سے زمین پر گر پڑے ، لوگوں نے دیکھا تومرغِ روح قفسِ عنصری سے پرواز کر چکا تھا۔ ’’انا ﷲ وانا الیہ راجعون ‘‘۔
مرحوم کی شخصیت عجیب وغریب کمالات واوصاف کی جامع تھی،حکومت کے اعلیٰ افسر ہونے کی حیثیت سے نہایت لائق وقابل، بڑے نیک نام اور حکومت اور پبلک دونوں کی نگاہ میں معتمد اورقابلِ احترام تھے۔ ہرمعاملہ میں سرکاری ہو یا غیرسرکاری اُن کی ایمان داری اوردیانت پرسب کاایمان تھا۔ ضرورت مندوں کے ساتھ ہمدردی وغمگساری اور عملاًان کی امداد واعانت...

پاکستان میں غیر مسلموں کے جان و مال کا تحفظ، قرآن وسنت کی روشنی میں خصوصی مطالعہ

This fact is quite obvious from Quran o Sunnah that Islam is a religion of peace and provides guarantee of protection of life, property and self respect to the all members of society without any discrimination of colour, race and religion. It is the duty of an Islamic State to provide protection to the basic rights of all minorities. Holy Prophetﷺ said: "Beware! Whoever is cruel and harsh to a non-muslims minority, curtailing their rights, overburdening them or stealing from them, I will complain (to God) about that person on the day of judgement." Islam seeks to establish such a society where all citizens of the state enjoy equal rights and religion doesn't become the basis from any discrimination. Islamic law holds both muslims and non-muslims equal and no superiority or privilege is given to the muslims on any ground.

Social Sector Organizations in Pakistan and Assessment of Their E- Accountability Practices

Social sector organizations can play an important role in meeting the development goals of a country; their true potential can be optimized if there is no trust deficit between all the stakeholders of the social sector. Social sector organizations should focus both on offline and online disclosures, according to international best practices; however, the importance of online disclosure carries more weight owing to the reliance of stakeholders on the internet. This study is an effort to start the debate pertaining to best practices about E-Accountability measures that social sector organizations should follow. For the case of Pakistan, there exist a schism between public, social sector organizations, and the government; the government of Pakistan revoked the licenses of many social sector organizations in 2016. It is important to note that positive contributions of social sector organization can be aptly highlighted if there is a third-party evaluation of their accountability mechanism. This study focused on the social sector organizations working in Pakistan; websites of 199 randomly sampled social sector organizations are analyzed by using a new E-Accountability index. Post-Stratification is used for the sampling of social sector organizations working in various service areas, such as education, and health. Due to the unavailability of a holistic directory of social sector organizations in Pakistan, we have developed a directory by using researching on multiple websites mentioned in detail in the main body of this study. The proposed E-Accountability index is defined according to three dimensions: usability, content, and communication, each of which contains different elements of downward, upward and functional accountability. The websites were coded in 2016. International social sector organizations performed better than the local ones. The contribution of print media is evident from the fact that those organizations that were featured in newspapers showed a better E-Accountability score. Size of the organization also played an important rule; social sector organizations with at least two branches score better on the E-Accountability index. Application of binary logistic regression on the proposed E-Accountability index showed that the odds of having a better E-Accountability score are 1.76 times higher for social sector organizations with branches. Odds of a better E-Accountability score for international social sector organizations are twice the odds for the local ones. The odds of a better E-Accountability score are 1.57 times higher for social sector organizations featured in newspapers as compared to the ones that are not featured. The proposed E-Accountability index can be used as a tool to issue ratings xx to social sector organization; these ratings can be further used by the government of Pakistan before issuing or renewing the license of these organizations. Better disclosures are observed about “Feedback Mechanism”, “Use of Other Media”, “List of Programs”, Clickability”, “Perceivabilty”, “Operability”, and link to “Social Networking Sites”. The worst disclosures are about “NGO Bylaws”, “Privacy Policy”, “Board of Directors Minutes”, Site in More than One Language”, “External Links to Local Organizations”, “Site Map”, Employee or Beneficiary Surveys”. The second contribution of this study is to evaluate a new model of social work i.e., Community Development Park Model, which is similar to themed science park except for the fact that instead of scientific discoveries these parks focus on solving social issues in an area by engaging multiple social sector organizations. Apparently, this seems unrelated to the proposed accountability framework; however, it must be kept in mind that the purpose of this research is to provide guidelines for best practices to SSOs working in Pakistan. Consequently, we presented the case of best practices followed by SSOs working in a Community Development Park Model. Rashidabad city in Tando Allahyar town of Sindh is using the Community Development Park Model to solve health, education, social inclusion, and employment issues of the local communities. Rashidabad offers an avenue of circumventing rural-urban integration; this park offers benefits of urban lifestyle with the serenity typical to rural areas. This park offers rural communities to feel connected with the urban lifestyles, hence shaping a common national identity. For comparison of education services, data were collected from 142 students enrolled at schools in Rashidbadad and 195 students enrolled at schools outside Rashidabad, 59 teachers from various schools within Rashidabad, and 28 teachers from various schools outside Rashidbabad. For comparison of health services, data were collected from 100 patients seeking health services from hospitals within Rashidabad, and 50 patients seeking health services from the District Headquarter Hospital, Tando Allahyar. Data collected from schools and hospitals inside and outside Rashidabad shows that schools and hospitals within Rashidabad outperform the ones outside Rashidabad. Within Rashidabad, perceptions of teachers are more positive, as compared to teachers of schools outside Rashidabad, about decision-making abilities of teachers, curriculum, collaboration between teachers & parents/students, financial resources, and human capital. Within Rashidabad, perceptions of students are more positive, as compared to students of schools outside Rashidabad, about quality of curriculum, exposure to IT skills, co-curricular activities, teaching standards, and medical-care. Assessment according to SAFED (2015) report card shows that the schools within Rashidabad possess all the facilities mentioned in xxi SAFED (2015) report i.e., usable toilet, safe drinking water, cleanliness of toilets, boundary wall, playground, computer lab, electricity connection, and library. Schools outside Rashidabad are worst in availability of playground, toilet cleanliness, and computer lab facility. Hospitals within Rashidabad are compared with District headquarter hospital, Tando Allahyar, using a total of 14 criterions: criterion 1 to 4 are about hospital staff, criterion 3 to 9 are about quality of arrangements for inpatients, criterion 10 to 11 are about pain management, criterion 12 to 14 are about hygiene & quality of food. Performance of District headquarter hospital remains invariably below the hospitals within Rashidabad; District headquarter hospital performs worst on the indicators pertaining to quality of food, theatre experience, staff identification, and bathroom facilities.