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Home > Detection of Mechanism of Invasive Activity of Burn Wound Pathogens and Their Susceptibility to Antibiotic and Natural Plant Products

Detection of Mechanism of Invasive Activity of Burn Wound Pathogens and Their Susceptibility to Antibiotic and Natural Plant Products

Thesis Info

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Author

Samad, Farkhunda

Program

PhD

Institute

University of Karachi

City

Karachi

Province

Sindh

Country

Pakistan

Thesis Completing Year

2017

Thesis Completion Status

Completed

Subject

Microbiology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10120/1/Farkhunda_Samad_Microbiology_SR_2017_UoK_karachi_18.07.2017.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725833996

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In emergency units burn injuries are considered to be the most agonizing injuries, ranging from minor to lethal. Developing countries such as Pakistan have a high incidence of burn injuries. Burn wound provides an ideal environment for the growth of pathogens. Burn wound serves as portal of invasion by microorganisms. Infections developed in a consequence of burn wound are a leading cause of death. Pseudomonas aeruginosa a non-spore forming, non-fermentative Gram negative bacillus, remains serious wound infection in burn patients and give rise to burn related morbidity and mortality worldwide. This study presents an overview about the mechanism of invasive activity of burn wound pathogens as well as ponders their alternative treatment options with natural plant products. To determine the spectrum of burn wound pathogens, a total of 350 pus samples were collected from patients having burn wound infections with total body surface area (TBSA) ranging from 1-90%. P. aeruginosa was observed in 26% cases as the main culprit of burn wound infection, irrespective of the type of thermal injuries. In addition, the incidence of bacteremia in burn patients was also determined in 150 patients and we observed bacteremia in 43(29%) of cases. It was important to note that in 15 cases the causative agent of bacteremia was P. aeruginosa which confirmed the well defined role of P. aeruginosa in burn patients. It was also of interest to evaluate time related changes in microbial colonization in burn wound infection. Fifty patients were followed up for four weeks at weekly interval. Results revealed that S. aureus was found to be the most frequently isolated pathogen during the 1st week while it was suppressed by Pseudomonas species from 2nd week. As for Candida spp., there was a gradual increase in the frequency of its recovery as time elapsed from admission. The risk regarding the acquisition of P. aeruginosa infection by burn patient is highly confronting because this organism is widely distributed in the hospital setting and can easily be transmitted to burn patients. To determine the exact role of the environment in the acquisition of P. aeruginosa by the burn patient, a total of 130 samples from different hospital sites including wall, floor, patient’s bed, Washing tubs and OT equipments was collected. It was interesting to find that in our results, distribution of P. aeruginosa varies with site of sample collection. Multidrug resistant bacteria have commonly been reported as colonizers of the wounds of burn patients. In order to select an effective antibiotic regimen to overcome the incidence of burn wound infection, the drug sensitivity pattern of burn wound pathogens were determined. Testing of 15 selected antibiotics yielded significant results, as much as 39% of the strains of S. aureus were found to be methicillin resistant. Vancomycin was the only “anti-staph” drug that inhibited the growth of all strains of S. aureus tested while linezolid acted on 95% of strains challenged. Additionally, polymyxin B and piperacillin/tazobactum were the drugs that found decidedly effective against Gram negative isolates of burn wound infection. The emergence of antimicrobial resistance among burn wound pathogens worldwide confine the possible therapeutic alternatives, used to treat burn wound infections. To ponder the alternative treatment options with natural plant products several plants were screened. Our finding reveled, extracts of Brassica rapa sub-spp. Campestris showed inhibitory activity against P. aeruginosa. On the basis of our observations extracts of Brassica rapa (SM2-PE and SM2-EA) were further investigated via gas chromatography-mass spectrometry (GC-MS) technique, for determination of bioactive components. 29 compounds were identified from SM2-PE and 35 compounds were identified from SM2-EA. The major classes of the identified compounds belonged to the monounsaturated fatty acid, methyl and ethyl esters. Both of the extracts were rich in fatty acid esters thus can be used as potential natural therapeutics. It is the first authenticated report on antibacterial activity and chemical composition ofBrassica rapa sub-spp. campestris from Pakistan. P. aeruginosa possessed several virulence factors such as, biofilm formation, exotoxin A and type III secretion system. These factors play important role in invasion, survival against the immunological responses of host and spread of infections. Biofilm formation ability of P. aeruginosa was detected by three methods. Among all tested P. aeruginosa isolates 57% were found to be biofilm producers. Additionally, genetic analysis of Exotoxin A gene sequences revealed the little difference in virulence gene distribution in P. aeruginosa isolated from different origins. 92% of P. aeruginosa from burn wound contained Exotoxin A genes while this gene was found in 89% of environmental strains of P. aeruginsa. Furthermore, prevalence oftype III secretion system gens (ExoS, ExoT, ExoU, ExoY) of P. aeruginosa from burn wound as well as environmental isolates were determined. Results revealed that all examined burn wound and environmental isolates harboured Exo T gene. While ExoU was found in 58% and 33% P. aeruginosa isolated from burn wound and environmental samples respectively. ExoS-was found in a minority of isolates. The prevalence of ExoSgenes varied from 36% in burn wound isolates and 6 % in environmental isolates. The PCR-based gene detection assay yielded that 90% of burn wound isolates and 10% of environmental isolates contained ExoY gene. To determine the expression of type III secretion exo-enzymes in the P. aeruginosa, we examined P. aeruginosa isolates from burn wound infection among them 5 were invasive strain (ExoS+/ExoU-) and 5 were cytotoxic strains (ExoS-/ExoU+) by (RT–PCR), under inducing condition all invasive strains of P. aeruginosa expressed ExoS while among cytotoxic strains only (40%) expressed ExoU. This is the first study from Pakistan providing evidence of a role for this type III secretion system in burn wound infection. On the basis of our observations, we conclude that burn wound infections and bacteremia are increasing in Pakistan. A continuous monitoring of burn wound pathogens as well as the up to date profiles of resistance to different drugs are necessary to develop better strategies for the treatment of burn patients. Biofilm forming property of P. aeruginosa not only contribute in increasing the severity of infection but it also participate in dilation of wound healing by effecting the defense mechanism of patient. P. aeruginosa that exhibited ETA gene may play an important role in the spread of infection. High prevalence of cytotoxic (ExoU+) strains of P. aeruginosa can induce severe inflammation because of its cytotoxic nature. Induced expression of ExoS proteins under depleted calcium conditions suggested that ExoS+ P. aeruginosa affects the immune system of patients which results in the accumulation of ExoS proteins in burn wound that can contribute in the development of infection. Type III secretion exoenzymes are a major virulence factor of P. aeruginosa which play important role in its pathogenicity, resulting in increased hospital stay and expense of treatment with significant economical burden.We are first time reporting that the fatty acid esters of Brassica rapa sub-spp. campestris Linn. can be use as potential natural therapeutics to treat not only burn wound infections but other infections as well. The results of this study will contribute in the development of new therapeutic agents to conquer the incidence of wound infections in burn patients and will benefit Pakistan economy by saving million of rupees, spend annually on patients who develop wound infection secondary to burn injury.
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۵۹۔ بابو

بابو٭

بابو!کس دیس چلا گیا تو

تیرے بعد اداس ہے بام کُو

نجانے کن پردوں کے پیچھے جا چھپا

کن ان دیکھی دیواروں کے پار اتر گیا

میری صدائیں ٹکڑا کر واپس پلٹ آتی ہیں

مگر تو نہیں آتا

کبھی دیوار سے جھانک تو سہی

تیرے بنا ہر پل ،ہر گھڑی

سرو و سمن اور کلی

چاند اور چاندنی سب اداس ہیں

جنگ، جہاد اور قتل

An elaborate endeavor in studying world religions would glaringly uncover the fact that most of the religions have enacted rules to follow, pertinent to daily livelihood. Such are the set of rules that the followers of the religion must follow. These rules very much include the rules about edibles. The food is something upon which not only human sustenance rests but it also keeps the human soul alive. For the same very reason we find religious and scientific prohibitions on things that are harmful for humans and if consumed evil takes the better of man’s heart, bringing upon him moral redundancy and sinister way of life. Such people are deprived of peace and piety. They ultimately not only suffer unbearable individual loss but also become a burden onto the society, country and nation

Comparativegrowthperformanceand Morphologicalvariationsanalysisbetween Two Populationsof Cirrhinus Mrigala from Chenab and Indus Rivers

In Pakistan there exist huge potential for development of aquaculture particularly inland fisheries sector but due to couple of reasons this field could not flourished in Pakistan. Cirrhinus mrigala is extensively cultured major carp among other major carps but there is no documentation concerning the analyses of Cirrhinus mrigala from Chenab River and Indus River for stock identification and conservation purposes. Length-weight and length-length relationships, condition factor, stock structure and morphological heterogeneity of Cirrhinus mrigala from Chenab River and Indus River were studied using traditional morphometric techniques; landmark (Truss) based geometric morphometric analysis and meristic analysis. A total of 248 samples were collected, 124 samples from each river. The higher value of regression coefficient ?b? from Indus river (b = 3.130) than regression coefficient ?b? value form Chenab River (b = 2.9875) indicated that Cirrhinus mrigala in Indus River shows comparatively better growth as compared to Chenab River. Length-weight relationships of Cirrhinus mrigala are suggestive that the wild conditions in Indus River at Taunsa Barrage are much suitable for rearing and stocking purposes instead in Chenab River at Head Muhammad Wala. Fulton?s condition factor (K) for Cirrhinus mrigala in Chenab River has shown a decreasing (negative) trend, while for the Indus River it showed an increasing (positive) trend. The higher evaluated level of Fulton?s condition factor for the population of Cirrhinus mrigala in the Indus River than the Chenab River population symbolizes the superior ecological conditions, foraging conditions, health and hygienic conditions in the Indus River at Taunsa Barrage. Discriminant function analysis on 21 landmark based geometric variables and 11 traditional morphometric variables disclosed that phenotypical variations are predominantly exist as Intra-population variations rather than inter-population variations. Final classification results in Discriminant function analysis, accounts that 67.7% of original grouped cases were correctly classified and 56.5% of cross-validated grouped cases were correctly classified. The equal discriminatory power of the discriminant function I (DFI) centred on discriminant function analysis for the both populations of Cirrhinus mrigala at Chenab River and the Indus River and non-significant (H) value of Kruskal-Wallis test patently represents the absence of statistically significant meristic variations between the two populations. Discriminant functional analysis and meristic analysis are of suggestive that there exists much merger and gene flow between two populations owing to weak geographical isolation between two rivers.