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Effect of Biocides on Biofilm Bacteria from Dental Plaque

Thesis Info

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Author

Saleem, Hafiz Ghulam Murtaza

Program

PhD

Institute

University of the Punjab

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2016-10

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/9270

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725954003

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The oral diseases (dental caries and periodontal diseases) are major public health problems and are found the most prevalent diseases of mankind. Accumulation and adherence of bacterial biofilm on tooth surfaces as dental plaque mainly cause for two human diseases like inflammatory periodontal disease and dental caries. The defensive immune systems and resident microbial flora are responsible for balance in oral health. Disruption of balance favours the colonisation of transient bacteria and commensal that may lead to localised infections e.g. periodontal, endodontic, and gingival infections. Oral cavity may provide suitable environment for the exchange and store of genetic material. Bacteria from the oral cavity have acquired resistance against antibiotics and/or biocides due to inadequate or extensive use of biocides and antibiotics in the field of dentistry and medicine. In this study five dental plaque samples were collected from five healthy individual for isolation, characterization and to check the efficacy of biocides on growth and biofilm formation of oral microbes. Susceptibility pattern of oral isolates were observed against different concentrations (0.1, 1, 10, 20, 30, 40, and 50 μg/ml) of four biocides/ commercially available mouth-washes by culture dependent method. These include Chlorhexidine (CHX) 0.2%, Benzidamine hydrochloride (BZD) 0.15%, Sodium fluoride 0.05% and potassium chloride 0.05%, and Benzidamine hydrochloride 0.15% and Chlorhexidine 0.2%). Thirty four (70%), 9 (18%), 2 (04%), 2 (04%) and 2 (04%) out of total 58 oral isolates were found with common biocides resistance pattern and showed reduce susceptibility against different concentrations 0.1, 1.0, 10, 20, 30 μgml-1of all four biocides respectively. All the 58 isolates were found sensitive at concentrations (40 and 50 μgml-1) of four biocides. Overall, 9 (15%) out of total fifty eight (58) plaque isolates were selected on different morphological basis and common resistance (1.0 μgml-1) against four biocides. Dental plaque bacteria were characterized morphologically, biochemically, physiologically and genetically; phylogenic analysis was also performed. The molecular characterization reveals that isolates N4A, B1E, N1D, N2C, N2D, N3C, N3D, N4E, and C2E belong to genera including Acinetobacter schindleri (JF837190), Morexiella acj (JF837191), Chryseobacterium culicis (KR002422), Chryseobacterium indologenes (KR002424), Acinetobacter johnsonii (KR002423), Enterobacter ludwigii (KR002425), Pseudomonas stutzeri (KC817808), Streptococcus salivarius (KC817808) and Bacillus cereus (JF837191) respectively. The isolates were variably resistant to multiple drugs including ampicillin, kanamycin, gentamicin and tetracycline, erythromycin, vancomycin and chloramphenicol. Seven out of nine isolates Summary 2 have shown resistance to ampicillin and tetracycline while erythromycin found the most effective drug with active susceptibility for six oral bacteria. Results indicated that all the plaque isolates were biofilm producers in the absence and presence of CHX within 24, 72 and 120 hours (hrs). High variation in biofilm forming ability of isolates was observed with exhibition of strong biofilm within 120 hrs followed by 72 hrs and 24 hrs. Study results directed that CHX+BZD with 1 μg/ml were found the most effective biocide for biofilm inhibition in 24 hrs. The effect of four biocides CHX, BZD, NaF+KCl and CHX+BZD (1.0 μg/ml stress) against biofilm bacteria in comparison with control (0 μg/ml stress) cells was analyzed statistically. It was found highly significant with P < 0.0001. Exposure of biofilms of these resistant species to undiluted commercial CHX mouthwash for intervals from 5 to 60 s indicated that the mouthwash was unlikely to eliminate them from dental plaque in vivo. Two species, Chryseobacterium indologenes and Chryseobacterium culicis were able to grow planktonically and form biofilms in the presence of 32 μg/ml CHX. Multidrug and CHX resistant C. indologenes demonstrated a 19-fold up-regulation of expression of gene CIN01S_RS05745 that encode the HlyD-like periplasmic adaptor protein of a tripartite efflux pump upon exposure to sub inhibitory concentration 16 μg/ ml of CHX suggesting that multidrug resistance may be mediated by this system. Bioinformatics analysis reveals the homology of this protein with RND family of transporters proteins (NCBI accession # TIGR01730) with e-value 2.08e-49. The signal of gene up regulation were also confirmed with the presence of 1250 bp amplification band of study gene flanking regions using reverse transcriptase PCR (RT-PCR). Northern hybridization reveals ̴6.3kb sized operon that encode for putative tripartite efflux pump. Furthermore, the ability of the selected actinomycetes to control biofilm of plaque bacteria was also accessed and observed that three out of ten Streptomyces strains named as S. erythrogriseus and S. labedae showed good antibacterial activity against Moraxella and Acinetobacter oral strains while S. macrosporeus can only inhibit biofilm formation by Bacillus cereus. Further study is required to show the transcriptomic picture of some CHX resistant genes in C. indologenes. This study has shown the resistance gene in emerging but uncommon pathogen; that will probably attract the researcher’s intension as new resistant bacterial species in dental setting. It has also pointed out the alarming situation with long term use of CHX (antiseptic/ oral care products) may cause resistance and/or may pose threat in the contribution of resistance gene to other dental microbiota. Future plan is to analyze the whole genome sequence data for genome comparison and for searching out CHX resistance efflux protein in newly Summary 3 sequenced C. culicis oral isolate. The whole genome sequencing data processing of C. culicis is going on for annotation using Rapid Annotation Subsystem Technology (RAST) tool.
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21. Al-Anbiya/The Prophets

I/We begin by the Blessed Name of Allah

The Immensely Merciful to all, The Infinitely Compassionate to everyone.

21:01
The time of their reckoning draws ever closer to the people as it starts while experiencing their death,
and yet in their heedlessness of its coming, they keep turning away.

21:02
Whenever a new Message comes to them from their Rabb - The Lord,
they listen to it playfully.

21:03
Their hearts are preoccupied with trivial things.
Yet, concealing their inner thoughts, the unjust/wrongdoers say in their private conversations:
‘Is this Muhammad not a human being like you?
Will you, then, submit to his magic, with your eyes wide open?’

21:04
He - The Prophet – said:
‘My Rabb - The Lord Knows whatever you confer and conspire and, in fact, every word
spoken within the celestial realm and the terrestrial world.
For HE is the All-Listening, the All-Knowing.’

21:05
No way!
‘Instead, they allege:
The Qur’an is just a collection of his jumbled false dreams.
Perhaps he has fabricated it.
Rather he is a poet!’
If this is not the case, then let him bring us a miraculous Sign, as were the former Messengers assigned with Signs.

21:06
Not one of the habitation(s) which WE destroyed in punishment before they had believed.
So, will they, then, be willing to believe or get destroyed?

21:07
And WE have not assigned any Messenger before you except for those men on to whom WE revealed -
so just ask scholars of the Former Divine Scriptures, if you do not know.

21:08
And WE did not make for...

طبي خدمات اور سہولیات کے تحفظ کي ضرورت: اسلامي شريعت کے چند رہنما اصول

Muslims scholars in principle agree that non-combatants are protected and that they lose protection when they directly participate in hostilities. However, the issues of defining the scope of non-combatant and that of direct participation remain contentious which resultantly cause confusions about the protection of medical personnel. The present paper digs out principles of Islamic law relating to the protection of medical personnel during armed conflict and for this purpose focuses on a doctor who works for humanity and who provides medical assistance to all and gives priority on the basis of need only. It tries to find answers to questions such as: is the doctor muqatil (combatant)? Does the act of providing medical assistance to the enemy combatants make the doctor liable for direct participation in hostilities? Does Islamic law distinguish between the legal consequences of direct and indirect participation in hostilities? After exploring the rich Islamic legal literature on the protection of medical personnel during armed conflict, the paper also examines the legal consequences of abuse of the protected status.

Synthesis and Characterization of Aliovalent Doped Bismuth Ferrite Multiferroics

Two batches of polycrystalline materials; Bi 1-x Ba x FeO 3 and Ba 1-2x Bi 2x Fe 1-x Nb x O 3 have been synthesized by solid state reaction method. The structural studies have been carried out by employing high resolution synchrotron X-Ray Diffraction (XRD), scanning electron microscopy (SEM) and Mössbauer spectroscopy. The dielectric properties are investigated by employing impedance spectroscopy. The DC electrical transport properties have been investigated by considering different conduction mechanisms. The magnetic properties have also been investigated at room temperature. The XRD data of the two batches have indicated a structural transition from rhombohedral to pseudo cubic symmetry at about 15% Ba substitution. The impurity phases are suppressed in the cation doped samples. The leakage current density in the Ba doped samples is reduced up to about four orders of magnitudes; however, the co- substitution with Ba 2+ and Nb 5+ in the compensated ratio has greatly reduced the leakage current up to about six order of magnitudes as compared to the undoped BiFeO 3 . The oxygen vacancies are found to have the dominant role in the leakage current. The overlapping large polaron tunneling (OLPT) and correlated barrier hopping (CBH) are the governing models for lightly doped samples in low temperature region and heavily doped samples at high temperatures, respectively. Although Nb doping is useful for the enhancement of the electrical resistivity; yet it significantly degrades the ferromagnetism appearing in the vicinity of Ba doping.