Text Figure 8 The dendrogram analysis

Transcription

1 Staphylococcus capitis subsp. capitis ATCC T (L37599) S. capitis subsp. ureolyticus ATCC T (AB009937) S. caprae ATCC T (AB009935) S. saccharolyticus ATCC T (L37602) 68 S. epidermidis ATCC T (D83363) S. aureus subsp. anaerobius ATCC35844 T (D83355) 100 SMKV SMKV- 2 S. aureus subsp. aureus ATCC12600 T (D83357) S. pasteuri ATCC51129 T AB S. lugdunensis ATCC T (AB009941) S. haemolyticus ATCC T (D83367) S. hominis subsp. hominis DSM T (X66101) S. kloosii ATCC T (AB009940) S. succinus subsp. succinus AMG-D1 T (AF004220) S. nepalensis CW1 T (AJ517414) 87 S. arlettae ATCC T (AB009933) S. cohnii subsp. cohnii ATCC29974 T (D83361) S. auricularis ATCC33753 T (D83358) S. chromogenes ATCC43764 T (D83360) S. felis ATCC49168 T (D83364) S. simulans ATCC T (D83373) S. piscifermentans ATCC51136 T (AB009943) S. carnosus ATCC T (AB009934) S. condimenti F-2 T (Y15750) S. sciuri subsp. sciuri ATCC T (S83569) 100 S. vitulus ATCC T (AB009946) Macrococcus caseolyticus ATCC13548 T (D83359) Text Figure 8 The dendrogram analysis

2 Percentage Text fig - 3 MRSA % prevalence in 2003, Southern region 90.00% 84.61% 80.00% 70.00% 69.56% 60.00% 53.12% 60.00% 61.29% 50.00% 48.14% 45.00% 40.00% 30.00% 31.50% 20.00% 10.00% 0.00% Age of the patient Below 1 55/65 1 to 10 16/23 11 to 20 13/27 21 to 30 17/32 31 to 40 18/30 41 to 50 19/31 51 to 60 9/20 61 to Above 6/19

3 PLATE - 1 PLATE - 2 MHA MRSA 1 Bacitracin 2 Doxycycline 3 Amikacin 4 Penicillin 5 Gentamycin MHA MRSA 1 Methicillin 2 Tetracycline 3 Ampicillin 4 Novobiocin PLATE - 3 MHA MSSA

4 PLATE - 4 PLATE - 5 Blood Agar - MRSA Mannitol Salt Agar - MRSA PLATE - 6 ELISA - MRSA C Control (showed blue colour) 1 6 MRSA inoculated (Absence of blue colour)

5 PLATE - 7 PLATE - 8 Rheumatoid arthritis Lesions in hind leg PLATE - 9 PLATE - 10 CONTROL MRSA Inoculated Liver shows White Patches

6 PLATE 11 Liver Control PLATE 12 Infected Liver 100 X 100 X PLATE 13 Kidney Control PLATE 14 Kidney Infected 100 X 100 X

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9 PLATE 22 Leaf of Azardicta indica juss PLATE 23 Leaf of Duranta pulmeneri jacq PLATE 24 Whole Plant of Punica granatum Linn.

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11 PLATE 30 -NMR

12 PLATE 29 - HPLC

13 PLATE 20 FIGE-REA MRSA Isolates TEXT FIGURE 7 Diagramatic Representation of FIGE-REA PLATE 21 mec A gene amplification

14 Plate 1: Staphylococcus aureus isolates grown on Muller- Hinton Agar plates showing its resistance against the antibiotics. (1) Bacitracin (2) Doxy cycline (3) Amikacin (4) Penicillin (5) Gentamycin Plate 2: Staphylococcus aureus isolates grown on Muller Minton Agar plates showing its resistance against as the antibiotics. (1) Methicillin (2) Tetracycline (3) Ampycillin (4) Novobiocin Plate 3: Plate showing methicillin sensitive ATCC Staphylococcus aureus strain No

15 Plate 4: Methicillin Resistant showing -haemolysin after 24 hrs incubation. Staphylococcus aureus grown on Blood Agar Plate 5: MRSA colonies grown on Mannitol Salt Agar, showing characteristic yellow colour. Plate 6: Test for the conformation of penicillinase production for MRSA (C) Control tube (1)-(5) MRSA isolates inoculated Absence of blue colour indicates the production of penicillinase.

16 Plate 7: MRSA infected mouse showing the characteristic symptoms for Rheumatoid Arthritis (RA). Plate 8: MRSA infected mice showing lesions in the hind leg. Plate 9: Dissected mice un inoculated with MRSA (control). Plate 10: Dissected mouse inoculated with MRSA. The Liver shows white patches.

17 Plate 11: Histopathological studies of normal liver observed under 100 X. Plate 12: Histopathology of infected liver observed under 100X showing characteristic necrosis in liver tissues. Plate 13: Histopathology of normal kidney cells taken from normal mice observed under 100 X. Plate 14: Histopathology of infected kidney cells from inoculated mice observed under 100X. Showing characteristic necrosis of kidney cells.

18 Plate 15: Protein profile of MRSA isolates on SDS- PAGE 1 = Standard Marker 2 = MRSA 1 3 = MRSA-2. 4 = MRSA-3. 5 = MRSA-4, 6 = MRSA-6 Plate 16: Rocket Immuno Electrophoresis of MRSA polyclonal antibodies raised in rabbits. Plate 17: Counter Immuno Electrophoresis of MRSA polyclonal Abs Raised in Rabbits. Precipitin arc formed in middle well.

19 Plate 18: Western Blot for MRSA cell wall proteins. Bands were showed at 5.8 kda & 22.5 kda region. Plate 19: Immunoblot for MRSA cell wall protein Bands showed at 5.8-kDa region.

20 Plate 20: FIGE REA of MRSA isolates. 1= Size Marker, 2=MTCC 87, 3= MRSA-1, 4=MRSA-2, 5=MRSA-3, 6=MRSA-4. Text figure 7: Diagrammatic representation of PFGE-FIGE-REA Plate 21: PCR amplification of mec A gene from MRSA isolates. Band obtained at 0.31-kbp region.

21 Plate 22: Leaf of Azardicta indica juss Plate 23: Leaf of Duranta pulmeneri jacq Plate 24: Whole plant of Punica granatum L

22 Plate 25: Plate showing the antibacterial activity of Neem & Duranta leaf extracts. Plate 26: Plate showing the antibacterial activity of Punica granatum L epicarp extract 1 = Tetracycline, 2 = Bacitracin, 3 = Novobiocin, 4 = Methicillin, 5 = Doxycycline, 6 = Ampicillin, 7 = Gentamycin, 8 = Extract Plate 27: Thin layer chromatography for Punica granatum L epicarp extracts. The sample showed 3 separate fractions as A, B & C. Plate 28: Plate showing antibacterial assay for the 3 fractions separated from TLC. 1 = Standard, 2 = Extract, 3 = Methicillin, 4 = Fraction A = Fraction B, 6 = Fraction C, 7 = Control.

23 Plate 29: High Performance Liquid Chromatography (HPLC) performed for the fractions A, B, & C obtained from TLC. Standard Peak obtained at Fraction A Peak obtained at Fraction B Peak obtained at Fraction C Peak obtained at 4.424

24 Plate 30: Hydrogen Nuclear Magnetic Resonance performed for the fractions A, B, C obtained from TLC. Fraction A, B & C produced peaks at the aromatic region (6 ppm).

25 LIST OF TEXT FIGURES TEXT FIGURE NO. TITLE 1 MRSA prevalence of southern region of Tamilnadu in the year of MRSA prevalence of southern region of Tamilnadu in the year of MRSA prevalence of southern region of Tamilnadu in the year of MRSA prevalence of northern region of Tamilnadu in the year of MRSA prevalence of northern region of Tamilnadu in the year of MRSA prevalence of southern region of Tamilnadu in the year of Diagrammatic representation of PFGE-FIGE-REA

26 LIST OF TABLES TABLE NO. TITLE 1 Clinical samples collected from southern region of Tamilnadu in the year S.aureus presented in southern region of Tamilnadu in the year MRSA prevalence southern region of Tamilnadu in the year Clinical samples collected from southern region of Tamilnadu in the year S.aureus presented in southern region of Tamilnadu in the year MRSA prevalence southern region of Tamilnadu in the year Clinical samples collected from southern region of Tamilnadu in the year S.aureus presented in southern region of Tamilnadu in the year MRSA prevalence southern region of Tamilnadu in the year Clinical samples collected from northern region of Tamilnadu in the year S.aureus presented in northern region of Tamilnadu in the year MRSA prevalence northern region of Tamilnadu in the year Clinical samples collected from northern region of Tamilnadu in the year S.aureus presented in northern region of Tamilnadu in the year MRSA prevalence northern region of Tamilnadu in the year Clinical samples collected from northern region of Tamilnadu in the year S.aureus presented in northern region of Tamilnadu in the year MRSA prevalence northern region of Tamilnadu in the year Percentage of MRSA prevalence of southern region of Tamilnadu in the year of Percentage of MRSA prevalence of southern region of Tamilnadu in the year of Percentage of MRSA prevalence of Northern region of Tamilnadu in the year of Percentage of MRSA prevalence of Southern region of Tamilnadu in the year of Percentage of MRSA prevalence of Northern region of Tamilnadu in the year of Percentage of MRSA prevalence of Northern region of Tamilnadu in the year of Protein estimation of MRSA cell wall protein. 14 Antibacterial activity of Azardicta indica juss, Duranta pulmeneri Jasq, punica granatum epicarp. 15 Rf value of Punica granatum epicarp fractions