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TITLE
DECLARATION
CERTIFICATE
ACKNOWLEDGEMENT
CONTENTS
ABBREVIATIONS
LIST OF FIGURES
LIST OF TABLES
1. INTRODUCTION
1 INTRODUCTION
1.1. AEROALLERGENS
1.2. SOURCES AND SlZE OF AEROALLERGENS
1.3. AIR SAMPLING
1.4. INTERPRETATION OF SAMPLING DATA FROM AIR SAMPLERS
1.5. ALLERGENIC FUNGI
1.6. CLASSES OF ALLERGENIC FUNGI
1.6.1. Oomycetes
1.6.2. Zygomycetes
1.6.3. Ascomycetes
1.6.4. Basidiomycetes
1.6.5. Deuteromycetes (Fungi imperfecti)
1.6.5.1. Sphaeropsidales
1.6.5.2. Melanoconoiales
1.6.5.3. Moniliales
1.7. ALLERGY
1.7.1. Types of allergy
1.7.2. Factors influencing allergenicity
1.8. IMMUNOGLOBULIN E (IgE)
1.8.1. IgE receptors
1.8.2. Allergen
1.9. ANTIGEN EXTRACTS
1.10. OBJECTIVES
2 REVIEW OF LITERATURE
2.1. AEROBIOLOGY
2.2. AEROMYCOLOGICAL STUDIES OUTSIDE INDIA
2.3. AEROMYCOLOGICAL STUDIES IN INDIA
2.4 CLINICO-IMMUNOLOGICAL STUDIES
2.4.1 Allergenicity of airborne fungal spores
2.4.2 Prevalence of fungal allergy
2.4.3 Manifestations
2. MATERIALS AND METHODS
2.1 AEROMYCOLOGICAL STUDY
2.2 INTRAMURAL SAMPLING SITES
2.3 SELECTION OF SAMPLING SITES
2.3.1 Bakery environment
2.3.2. Vegetable market
2.3.3. Hospital ward
2.3.4. Patients bedroom
2.4. SAMPLING rECHNIQUES
2.4.1. Air sampler
Fig. 1 Six stage Anderson type sampler with vacuum pump.
Fig. 2 Anderson type sampler with six stages shown separately along with vacuum pump.
1. Six stage Anderson type sampler with vacuum pump.
2. Anderson type sampler with six stages shown separately along with vacuum pump.
2.4.2. Sampling procedure
2.5. ISOLATION AND IDENTIFICATION OF AIRBORNE FUNGI
2.6. EXPRESSION OF AEROMYCOLOGICAL DATA
2.6.1. Calculation
2.6.2. Dominant and frequent fungi
2.7. CLINICO-IMMUNOLOGICAL METHODS
2.8. COLLECTION AND STORAGE OF FUNGI
2.9. MASS CULTURING OF THE ISOLATED FUNGI
2.10. HARVESTING AND DRYING
2.11. GRINDING
2.12. DEFATTING
2.13. PREPARATlON OF FUNGAL ANTIGEN
2.13.1. Extraction
2.13.2. Clarification
2.13.3. Sterilization
2.14. STERlLlTY TESTING
2.15. STANDARDIZATION
2.16. SKIN TEST
2.16.1 Intradermal testing procedure of asthmatic patientswith the extracted fungal antigens (~hivfiri, 1962)
2.16.2 Definition and grading of positive intradermal skin testreaction (Shivpuri, 1962)
2.16.3. Patient selection for allergy evaluation
2.16.4. Inclusion criteria
2.16.5. Exclusion criteria
2.16.6. Free informed consent
2.16.7. Freedom from assault
2.16.8. Confidentiality
2.17. STATISTICAL METHODS
2.17.1 Explanation of Tests
2.17.2. Regression Analysis
3. RESULTS
3.1. AEROMYCOLOGICAL STUDIES
3.2. FUNGAL TYPES
3.2.1 AspergilIus niger
3. Aspergillus niger. LM picture of conidiophore and conidia: X400.
4. Aspergillus roger. SEM picture of conidia: X9000.
5. Aspergillus niger. SEM picture of conidiophore and conidia: X1300.
6. Aspergillus flavus. LM picture of conidiophore and conidia: X400.
7. Aspergillus flavus. SEM picture of conidiophore and conidia: X1500.
8. Aspergillus flavus. SEM picture of conidia: X12000.
9. Aspergillus fumigatus. LM picture of conidiophore and conidia: X400.
10. Aspergillus fumigatus. SEM picture of conidiophore and conidia: X1000.
3.2.2 Aspergillus Flavus
3.2.3 Aspergillus fumigatus
3.2.4 Penicillium funiculosum
11. Aspergillus fumigatus. SEM picture of conidia: X2000.
12. Penicillium funiculosum LM picture of Penicilli and conidia: X400.
13. Pen icillium funiculosum. SEM picture of Penicillus and conidia: X4000.
14. Penicillium oxalicum. LM picture of Penicilli and conidia: X400.
15. Penicillium oxalicum. SEM Picture of Phialide and conidia: X1000.
16. Penicillium citrinum. LM picture of Penicilli and conidia: X400.
17. Penicillium citrinum. LM picture of Phialide and conidia: X1000.
18. Penicillium expansum. LM picture of Penicilli and conidia: X400
3.2.5. Penicillium oxalicum
3.2.6. Penicillium citrinum
3.2.7. penicillium expansum
3.2.8. Penicillium chrysogenum
19. Penicillium expansum. SEM picture of Penicilli and conidia: X1500.
20. Penicillium chrysogenum SEM picture of Penicilli and conidia: X3500.
21. Cladosporium cladosporioides. LM picture of conidiophore and conidia: X1000.
22. Cladosporium cladosporioides. SEM picture of condiophore and conidia: X2000.
23. Alternaria alternata. LM picture of conidiophore and condia: X1000
24. Rhizopus stolonifer. LM picture of sporangium and sporangiospores: X400.
25. Rhizopus stolonifer. SEM picture of sporangiophore, collumella and sporangiospores: X2000.
26. Rhizopus stolonifer. Sporangiospore: X8500
3.2.9. Cladosporiurn cladosporioides
3.2.10. Alternuria alternata
27. Candida albicans SEM picture of blastospores: X10000
28. Fusarium oxysporum. LM picture of macroconidia and microconidia: X1000.
3.2.11. Rhizopus stolonifer
3.2.12. Cundida albicans
3.2.13. Fusarium oxysporum
3.3. PREVALENCE OF ATMOSPHERIC FUNGAL TYPES
3.4. BAKERY ENVIRONMENT
3.4.1. Monthly average fungal concentration of dominant and frequentfungal concentration (CFU/m3) inside and outside bakeryenvironment
1. Monthly average of dominant and frequent fungal concentration (CFU / m3) inside bakery environment during November 1998 - October 2000.
2. Monthly average of dominant and frequent fungi concentration (CFU / m3) outside bakery environment during November 1998 - October 2000.
3.4.2. Annual average colony concentration and percentage contributionof dominant and frequent fungal concentration (CFUm3) inside and outside bakery environment
3. Colony concentration and percentage contribution of fungal types inside and outside bakery.
3.4.3. Statistical significance in the comparison of means of dominantfungal colony concentration inside and outside bakeryenvironment i
4. Statistical significance of dominant colony concentration inside and outside bakery.
3.4.4. Seasonal variation of dominant fungal concentration (c17u/m3) inside and outside bakery environment
5. Seasonal variation of Aspergillus niger inside and outside bakery.
6. Seasonal variation of Aspergillus flavus inside and outside bakery.
7. Seasonal variation of Cladosporium spp. inside and outside bakery.
8. Seasonal variation of Penicillium oxalicum inside and outside bakery.
9. Seasonal variation of Alternaria spp. inside and outside bakery.
3.4.5. Effect of temperature (T) on total dominant colony concentration (cFu/~) inside and outside bakery environment.
10. Influence of temperature (T) on total dominant fungi concentration (CFU / m3) inside and outside bakery.
3.4.6. Effect of relative humidity (RH) on total dominant colonyconcentration (C F U I ~in~sid e and outside bakery environment
11. Regression analysis of dominant fungal colony concentration in bakery with temperature.
12. Influence of relative humidity (RH) on total dominant fungal concentration (CFU / m3) inside and outside bakery.
13. Regression analysis of dominant colony concentration in bakery with relative humidity.
3.5. VEGETABLE MARKET ENVIRONMENT
3.5.1. The monthly average fungal concentration (c F u / ~ ~i) n side andoutside vegetable market environment
14. Monthly average of dominant and frequent fungal concentration (CFU / m3) inside vegetable market environment during November 1998 - October 2000.
15. Monthly average of dominant and frequent fungal concentration (CFU/ m3) outside vegetable market during November 1998 - October 2000.
3.5.2. Average annual colony concentration and percentage contributionof dominant and frequent fungal concentration (C F U I ~ ~in) si deand outside vegetable market environment
16. Colony concentration and percentage contribution of fungal types inside and outside vegetable market.
3.5.3. Statistical significance in the comparison of means of dominantfungal concentration inside and outside vegetable marketenvironment
3.5.4. Seasonal variation of dominant fungal concentration inside andoutside vegetable market environment
17. Statistical significance of dominant colony concentration inside and outside vegetable market.
18. Seasonal variation of Cladosporium spp. inside and outside vegetable market.
19. Seasonal variation of Aspergillus niger inside and outside vegetable market.
20. Seasonal variation of Aspergillus flavus inside and outside vegetable market.
21. Seasonal variation of Penicillium citrinum inside and outside vegetable market.
22. Seasonal variation of Pencillium oxalicum inside and outside vegetable market.
3.5.5. Effect of temperature (T) on total dominant colony concentration (C F U I ~in~s) id e and outside vegetable market environment
23. Influence of temperature (T) on total dominant fungal concentration (CFU/ m3) inside and outside vegetable market.
3.5.6. Effect of relative humidity (RH) on total dominant colonyconcentration (c F u I ~ ~in~s) id e an8 outside vegetable marketenvironment.
24. Regression analysis of dominant colony concentration of vegetable market with temperature.
25. Influence of relative humidity (RH) on total dominant fungal concentration (CFU/ m) inside and outside vegetable market.
3.6. HOSPITAL WARD ENVIRONMENT
26. Regression analysis of dominant colony concentration of vegetable market with relative humidity.
3.6.1. The monthly average fungal concentration inside and outsidehospital ward environment.
27. Monthly average of dominant and frequent fungal concentration (CFU/ m3) inside hospital ward environment during November 1998 - October 2000.
28. Monthly average of dominant and frequent fungal concentration (CFU/ m3) outside hospital ward during November 1998 - October 2000.
3.6.2. Annual average colony concentration and percentage contributionof dominant and frequent fungal concentration (CFU/m3) insideand outside hospital ward environment
29. Colony concentration and percentage contribution of fungal types inside and outside hospital ward.
3.6.3. Statistical significance in the comparison of means of dominantcolony concentration inside and outside hospital ward
3.6.4. Seasonal variation of dominant fungal concentration (CFU/m3) inside and outside hospital ward
30. Statistical significance of dominant colony concentration inside and outside hospital ward.
31. Seasonal variation shown by Aspergillus niger inside and outside hospital ward.
32. Seasonal variation of Cladosporium spp. inside and outside hospital ward.
33. Seasonal variation of Penicillium oxalicum inside and outside hospital ward.
34. Seasonal variation of Aspergillus flavus inside and outside hospital ward.
3.6.5. Effect of temperature (T) on total dominant colony concentration (CFU/m3) inside and outside hospital ward
35. Influence of temperature (T) on total dominant fungi concentration (CFU/ m) inside and outside hospital ward.
3.6.6. Effect of relative humidity (RH) on total dominant colonyconcentration (CFU/m3) inside and outside hospital ward
36. Regression analysis of dominant colony concentration of hospital ward with temperature.
37. Influence of relative humidity (RH) on total dominant fungi concentration (CFU/ m3) inside and outside hospital ward.
38. Regression analysis of dominant colony concentration of hospital ward with relative humidity.
3.7. ASTHMATIC PATIENTS BEDROOM ENVIRONMENT
3.7.1. The monthly average fungal concentration inside asthmaticpatients bedroom and outside
39. Monthly average of dominant and frequent fungal concentration (CFU/ m3) inside asthmatic patients bedroom environment during November 1998 - October 2000.
40. Monthly average off dominant and frequent fungal concentration (CFU/ m3) outside asthmatic patients bedroom environment during November 1998 - October 2000.
3.7.2. Annual average colony concentration and percentage contributionof dominant and frequent fungal concentration (C F U I ~ ~in) si deasthmatic patients bedroom and outside
41. Colony concentration and percentage contribution of fungal types inside asthmatic patients bedroom and outside.
3.7.3. Statistical Significance in the comparison of means of dominantfungal colony concentration inside asthmatic patients bedroomand outside
3.7.4. Seasonal average variation of dominant fungal concentration (CFU/m3) inside asthmatic patients bedroom and outside
42. Statistical significance of dominant fungi colony concentration inside asthmatic patients bedroom and outside.
43. Seasonal variation of Aspergillus niger inside asthmatic patients bedroom and outside.
44. Seasonal variation of Cladosporium spp.inside asthmatic patients bedroom and outside.
45. Seasonal variation of Penicillium citrinum inside asthmatic patients bedroom and outside.
46. Seasonal variation of Aspergillus fumigatus inside asthmatic patientsbedroom and outside.
47. Seasonal variation of Penicillium oxalicum inside and outside asthmatic patients bedroom.
3.7.4. Effect of temperature (T) on total dominant colony concentration (CFUI~) inside asthmatic patients bedroom and outside.
48. Seasonal variation of Penicillium expansum inside asthmatic patients bedroom and outside.
49. Influence of temperature (T) on total dominant colony concentration (CFU/ m3) inside asthmatic patients bedroom and outside.
50. Regression analysis of dominant colony concentration inside asthmatic patients bedroom and outside with temperature.
3.7.5. Effect of relative humidity (RH) on total dominant colonyconcentration (C: FUI~) inside asthmatic patients bedroom andoutsid
51. Influence of relative humidity (RH) on total dominant colony concentration (CFU/ m3) inside asthmatic patients bedroom and outside.
52. Statistical significance of dominant colony concentration inside asthmatic patients bedroom and outside with relative humidity.
3.8. CLINIC0 - IMMUNOIAOGICAL STUDIES
3.8.1. RESULTS OF SKIN TESTS
3.8.2. SEXWISE CI.ASSIF1CATION OF PATIENTS IN DIFFERENT AGE
53. Age groups and sex of patients studied.
3.8.3. REACTION TO Penicillium oxalicum
54. Agewise classification of reaction to Penicillium oxalicum.
3.8.4. REACTIONS TO Alternaria spp.
55. Sexwise classifcation of reaction to Penicillium oxalicum.
56. Agewise classification of reaction to Alternaria spp.
57. Sexwise classification of reaction to Alternaria spp.
3.8.5. REACTION TO Penicifiium expansum
58. Agewise classification of reaction to Penicillium expansum
3.8.6. REACTION TO Rhizopus stolonifer
59. Sexwise classification of reaction to Penicillium expansum.
60. Agewise classification of reaction to Rhizopus stolonifer.
61. Sexwise classification of reaction to Rhizopus stolonifer.
3.8.7. REACTION TO Penicilliurn citrinurn
62. Agewise classification of reaction to Penicillium citrinum.
3.8.8. REACTION TO Candida afbicans
63. Sexwise classification of reaction to Penicillium citrinum.
64. Agewise classification of reaction to Candida albicans.
65. Sexwise classification of reaction to Candida albicans.
3.8.9. REACTION TO Aspergillus furnigatus
66. Agewise classification of reaction to Aspergillus fumigatus.
3.8.10. REACTION TO Cladosporium spp.
67. Sexwise classification of reaction to Aspergillus fumigatus.
68. Agewise classification of reaction to Cladosporium spp.
3.8.11. REACTION TO Aspergillusflavus
69. Sexwise classification of reaction to Cladosporium spp.
70. Agewise classification of reaction to Aspergillus flavus.
71. Sexwise classification of reaction to Aspergillus flavus.
3.8.12. REACTION TO Asprugillus niger
72. Agewise classification of reaction to Aspergillus niger.
3.8.13.COMPARlSON OF THE PERCENTAGS OF SKIN TESTREACTIONS WITH CLINICALLY SlGNIFICANT ANTIGENS
73. Sexwise classification of reaction to Aspergillus niger.
74. Comparison of the skin test reactions with antigens.
3.8.14. CLINICALLY SIGNIFICANT FUNGI WITH SKIN REACTIONS 2+AND ABOVE
75. Fungi with 2+ and above skin reactions.
4. DISCUSSION
4.1. AIR BORNE FUNGI IN BAKERY
4.1.1. Colony concentration and percentage contribution of dominantand frequent fungi inside and outside bakery
4.1.2. Seasonal variation
4.1.3. Temperature
4.1.4. Relative humidity (RH)
4.2. AIR BORNE FUNGI IN VEGETABLE MARKET
4.2.1 Colony concentration and percentage contribution of dominantfungi inside and outside vegetable market
4.2.2. Seasonal variation
4.2.3. Temperature
4.2.4. Relative humidily (RH)
4.3. AIR BORNE FUNGl IN HOSPITAL WARD
4.3.1. Colony concentration and percentage contribution of dominantand frequent fungi inside and outside hospital ward
4.3.2. Seasonal variation
4.3.3. Temperature
4.3.4. Relative humidity (KH)
4.4. AIR BORNE FUNGI IN ASTHMATIC PATIENTS BEDROOM
4.4.1. Colony concentration and percentage contribution of dominantand frequent fungi inside asthmatic patientsbedroom andoutside
4.4.2. Seasonal variation
4.4.3. Temperature
4.4.4. Relative humidity (RH)
4.5. CLINICO - IMMUNOLOGICAL STUDIES
4.6. SKIN TESTS
4.6.1. Agewise and sexwise skin test postivity of asthmatic patients toclinically significant antigens
4.6.2. Skin test postivity of asthmatic patients to - clinically significantantigens
4.7. SIGNIFICANCE OF THE PRESENT STUDY
4.8. SCOPE FOR FUTURE WORK
5. SUMMARY AND CONCLUSION
REFERENCES
APPENDIX
SITE 1: BAKERY ENVIRONMENT
SITE 2: VEGETABLE MARKET ENVIRONMENT
SITE 3: HOSPITAL WARD ENVIRONMENT
SITE 4: ASTHMATIC PATIENTS BEDROOM ENVIRONMENT