1. Manjunath KManjunath K
Chairperson, Department ofChairperson, Department of
Microbiology and Biotechnology,Microbiology and Biotechnology,
Bangalore UniversityBangalore University
The Relentless Rise ofThe Relentless Rise of
Neoallergens inNeoallergens in
EnvironmentEnvironment

2. HISTORYHISTORY
► Foundation of Allergy, i.e. “Disorders of ImmuneFoundation of Allergy, i.e. “Disorders of Immune
System where the balance is lost resulting inSystem where the balance is lost resulting in
morbidity and mortality” started withmorbidity and mortality” started with
►1873 Chails blackley explained pollen as1873 Chails blackley explained pollen as
causative agent.causative agent.
►1919 John Bostock described Hayfever1919 John Bostock described Hayfever
►1921 Prausnitz Kurtner discovered IgE in1921 Prausnitz Kurtner discovered IgE in
serum.serum.
►1967 Johansen suggested the name “Ig E”1967 Johansen suggested the name “Ig E”

3. ► In the last 30 yrs spectacular development inIn the last 30 yrs spectacular development in
cellular and molecular mechanism of allergiccellular and molecular mechanism of allergic
diseases has been done.diseases has been done.
► In the new millennium almost half theIn the new millennium almost half the
populations of the West show sensitization topopulations of the West show sensitization to
one or other environmental allergensone or other environmental allergens
► Britain: one in four children has AsthmaBritain: one in four children has Asthma
► Australia: one in five has EczemaAustralia: one in five has Eczema
► People are found allergic to new allergensPeople are found allergic to new allergens
like nuts, soya, latex etc.like nuts, soya, latex etc.

4. ORGANIC DUSTORGANIC DUST► Genera grain dustGenera grain dust
► Wheat/Rice/Ragi/Jowar dustWheat/Rice/Ragi/Jowar dust
► Soya dustSoya dust
► Cotton dustCotton dust
► KapokKapok
► FlexFlex
► HempHemp
► JuteJute
► StrawStraw
► SilkSilk
► Castor bean dustCastor bean dust
► Coffee bean dustCoffee bean dust
► Tea dustTea dust
► Rye/ oat dustRye/ oat dust
► House dust (MITES)House dust (MITES)
► Sick buildings (Damp walls) Dander/feathers ofSick buildings (Damp walls) Dander/feathers of
pets/sheep/cattle/poultry/pigs/cockroaches/ratspets/sheep/cattle/poultry/pigs/cockroaches/rats
► Palmolein oilPalmolein oil

5. OCCUPATIONAL ENVIRONMENTSOCCUPATIONAL ENVIRONMENTS
► Leather Godowns and industryLeather Godowns and industry
► Flour mill/BakeryFlour mill/Bakery
► Sugarcane/Brewery/SCP factory (spirulina)Sugarcane/Brewery/SCP factory (spirulina)
► Bioinsecticide industry like Bt toxinBioinsecticide industry like Bt toxin
► Mushroom factoryMushroom factory
► Sewage treatment plantsSewage treatment plants
► Viral/Bacterial vaccine production unitsViral/Bacterial vaccine production units
► composting and garbage handlingcomposting and garbage handling
► fiber glass productionfiber glass production
► metal machiningmetal machining
► dental clinicsdental clinics

6. OCCUPATIONALOCCUPATIONAL
ENVIRONMENTSENVIRONMENTS
►PoultryPoultry
►Swine confinementSwine confinement
►Dairy barns and Livestock productionDairy barns and Livestock production
►Grain handling facilitiesGrain handling facilities
►cigarette and cigar manufacturingcigarette and cigar manufacturing
►animal feed industryanimal feed industry
►potato processingpotato processing
►paper factorypaper factory

7. ►sawmills and wood working shopssawmills and wood working shops
►cotton processing, jute mill, gin housecotton processing, jute mill, gin house
►BiotechnologyBiotechnology
►OfficesOffices
►HousesHouses
OCCUPATIONALOCCUPATIONAL
ENVIRONMENTSENVIRONMENTS

8. AIRBORNE BACTERIA /AIRBORNE BACTERIA /
ENDOTOXINENDOTOXIN
► Airborne BacteriaAirborne Bacteria varies from 10varies from 1055
-10-1099
CFU/mCFU/m33
in agriculturalin agricultural
and industrial environmentsand industrial environments
► In general terms, ‘endotoxins’ refers to any cell-boundIn general terms, ‘endotoxins’ refers to any cell-bound
bacterial toxin - a characteristic that distinguishes it frombacterial toxin - a characteristic that distinguishes it from
‘exotoxins’ which are secreted by bacterial cells‘exotoxins’ which are secreted by bacterial cells
► Heteropolymers- polysaccharides- immunologic + lipid A - toxicHeteropolymers- polysaccharides- immunologic + lipid A - toxic
► Released as discoid particles 30-50nm diameter in largeReleased as discoid particles 30-50nm diameter in large
quantity into airquantity into air
► Endotoxin + Lipopolysaccharide binding protein (LBP) via CD4Endotoxin + Lipopolysaccharide binding protein (LBP) via CD4
receptor alveolar macrophage binding site and activate immunereceptor alveolar macrophage binding site and activate immune
systemsystem
► E.coli, Salmonella, Shigella,Pseudomonas, Haemophilus,E.coli, Salmonella, Shigella,Pseudomonas, Haemophilus,
NeisseriaNeisseria
► Airborne bacteria urban site 609 CFU/mAirborne bacteria urban site 609 CFU/m33
(Lightart 1997)(Lightart 1997)
► Bacteria increase sunrise decrease noon increase sunsetBacteria increase sunrise decrease noon increase sunset
decrease eveningdecrease evening

9. Antibiotic Resistant bacteria in Indoor Air
► In Indoor air the most important source of airborne bacteria is human activity
► Extended-spectrum cephalosporin-resistant Gram-negative bacilli
► MDR Pseudomonas aeruginosa and carbepenem resistant Acinetobacter.
► Methicillin Resistant Staphylococcus aureus and Methicillin Resistant Coagulase
negative Staphylococci
► Health risk associated with occupant exposure to airborne bacteria are difficult
to calculate because of fewer studies.
► Increased mortality and hospital costs
► More surveillance data is required from India to frame
national guidelines and policies to combat antibiotic
resistance

10. Table:1Table:1 OCCUPATIONAL EXPOSURE TOOCCUPATIONAL EXPOSURE TO
AIRBORNE BACTERIA AND ENDOTOXINAIRBORNE BACTERIA AND ENDOTOXIN
(Agricultural Industry)(Agricultural Industry)
SiteSite
Concentration ofConcentration of
bacteria in the airbacteria in the air
(CFU x 10(CFU x 10 33
/m/m 33
))
RespirableRespirable
fraction (%)fraction (%)
Concentration ofConcentration of
endotoxin in the airendotoxin in the air
(CFU x 10(CFU x 10 33
/m/m 33
))
Cow barnsCow barns
Horse stablesHorse stables
PiggeriesPiggeries
Poultry housesPoultry houses
Silo unloadingSilo unloading
Grain HandlingGrain Handling
43.4-282.143.4-282.1
27.5-308.527.5-308.5
300.0-8,000.0300.0-8,000.0
41.2-7596.541.2-7596.5
60.0-4,300,000.060.0-4,300,000.0
70.0-1.244.870.0-1.244.8
51.251.2
40.040.0
44.844.8
57.857.8
38.138.1
41.9-54.941.9-54.9
0.0012-0.0120.0012-0.012
0.006-3.440.006-3.44
0.0019-75.00.0019-75.0
0.13-1.420.13-1.42
0.16-8.850.16-8.85
0.1-54.90.1-54.9

11. Table:2Table:2 OCCUPATIONAL EXPOSURE TOOCCUPATIONAL EXPOSURE TO
AIRBORNE BACTERIA AND ENDOTOXINAIRBORNE BACTERIA AND ENDOTOXIN
SiteSite
Concentration ofConcentration of
bacteria in the airbacteria in the air
(CFU x 10(CFU x 10 33
/m/m 33
))
RespirableRespirable
fraction (%)fraction (%)
Concentration ofConcentration of
endotoxin in the airendotoxin in the air
(CFU x 10(CFU x 10 33
/m/m 33
))
Poultry processingPoultry processing
Animal FeedAnimal Feed
industryindustry
BreweriesBreweries
Vegetable SeedVegetable Seed
storesstores
HerbageHerbage
processingprocessing
Slicing beet slicingSlicing beet slicing
29.6-1,273.929.6-1,273.9
161.7-1,452.4161.7-1,452.4
23.7-526.623.7-526.6
10.9-8,991.410.9-8,991.4
36.1-518.236.1-518.2
179.1-203.3179.1-203.3
33.333.3
Not TestedNot Tested
Not TestedNot Tested
41.141.1
44.344.3
Not TestedNot Tested
0.63-0.920.63-0.92
0.0250.025
0.06-0.930.06-0.93
Not TestedNot Tested
0.18-2681.00.18-2681.0
0.003-0.0320.003-0.032

12. Table:3Table:3 OCCUPATIONAL EXPOSURE TOOCCUPATIONAL EXPOSURE TO
AIRBORNE BACTERIA AND ENDOTOXINAIRBORNE BACTERIA AND ENDOTOXIN
SiteSite
Concentration ofConcentration of
bacteria in the airbacteria in the air
(CFU x 10(CFU x 10 33
/m/m 33
))
RespirableRespirable
fraction (%)fraction (%)
Concentration ofConcentration of
endotoxin in the airendotoxin in the air
(CFU x 10(CFU x 10 33
/m/m 33
))
Cotton millsCotton mills
Jute millsJute mills
Sawing woodSawing wood
Chipboard factoriesChipboard factories
EngineeringEngineering
IndustryIndustry
Composing wasteComposing waste
RecyclingRecycling
17.5 - 1,285.917.5 - 1,285.9
109.1-1,143.9109.1-1,143.9
5.7-37.05.7-37.0
21.4-36.321.4-36.3
0.2-735,135.10.2-735,135.1
100.0- 30,645.0100.0- 30,645.0
4.7-54.44.7-54.4
Not TestedNot Tested
Not TestedNot Tested
20.4-37.020.4-37.0
35.4-63.235.4-63.2
Not TestedNot Tested
50-6050-60
Not TestedNot Tested
0.001-2.20.001-2.2
0.02-0.050.02-0.05
0.023-4.00.023-4.0
1.23-40.01.23-40.0
Not TestedNot Tested
0.001-0.040.001-0.04
0.0008-0.00250.0008-0.0025

13. Categories of CFU/m3
(mixed population of bacteria and
fungi) for non-industrial indoor environments (CEC, 1993)
Category Bacteria Fungi
Very Low <50 <25
Low <100 <100
Intermediate <500 <500
High <2000 <2000
Very High >2000 >2000

14. Proposed Occupational Exposure
Limits (OEL) values
Malmros et al. 1992 Total airborne
microorganisms
10 × 103
cfu/m3
Dutkiewicz and
Jablonski, 1989
Total airborne
microorganisms
100 × 103
cfu/m3
Clark, 1985
Malmros et al.1992
Gram-negative bacilli 1 × 103
cfu/m3
Dutkiewicz and
Jablonski, 1989
Gram-negative bacilli 20 × 103
cfu/m3

15. SEWAGE PLANTSSEWAGE PLANTS
►Endotoxins are estimated by LAL TESTEndotoxins are estimated by LAL TEST
►BACTERIA 105 CFU/mBACTERIA 105 CFU/m33
►ENDOTOXIN 410ng/mENDOTOXIN 410ng/m33
►STANDARD LIMIT OF EXPOSURE 30-50STANDARD LIMIT OF EXPOSURE 30-50
ng/m3ng/m3
►Bacterial count on Eosin Methylene BlueBacterial count on Eosin Methylene Blue
►Agar 5400 CFU/mAgar 5400 CFU/m33
►R2A Agar 34500 CFU/mR2A Agar 34500 CFU/m33

16. TOXINS AS ALLERGENSTOXINS AS ALLERGENS
1. Streptococcal toxin1. Streptococcal toxin
2. Staphylococcal enterotoxin2. Staphylococcal enterotoxin
3. Aeromonas hydrophilic enterotoxin3. Aeromonas hydrophilic enterotoxin
These act as superantigensThese act as superantigens
Directly induce immune response bypassing usual processDirectly induce immune response bypassing usual process
of peptide presentation toof peptide presentation to αβαβ chain of T cells. Insteadchain of T cells. Instead
directly react with surface of IgG on B cellsdirectly react with surface of IgG on B cells..
4. Aflatoxin4. Aflatoxin
5.Mycotoxins5.Mycotoxins
6. Phycotoxins6. Phycotoxins

17. EFFECTS OF ENDOTOXINSEFFECTS OF ENDOTOXINS
EXPOSURE 100-200 EU /mEXPOSURE 100-200 EU /m33
1.1. AIRWAYS INFLAMMATIONAIRWAYS INFLAMMATION
2.2. HYPERSENSITIVITY PNEUMONITIS, ALLERGICHYPERSENSITIVITY PNEUMONITIS, ALLERGIC
ASTHMAASTHMA
3.3. LUNG INJURYLUNG INJURY
4.4. ABDOMINAL CRAMPS, DIARRHOEA,ABDOMINAL CRAMPS, DIARRHOEA,
5.5. DRY COUGH, BREATHLESSNESSDRY COUGH, BREATHLESSNESS
6.6. SNEEZINGSNEEZING
7.7. ADJUVANT EFFECTSADJUVANT EFFECTS
8.8. SYSTEMIC EFFECTSSYSTEMIC EFFECTS
9.9. CHRONIC BRONCHITISCHRONIC BRONCHITIS
10.10. CHEST TIGHTNESS, WHEEZINGCHEST TIGHTNESS, WHEEZING

18. Endotoxin activity

19. AIR SAMPLERS AND RECENTAIR SAMPLERS AND RECENT
TECHNIQUESTECHNIQUES
1.1. DURHAM SAMPLERDURHAM SAMPLER
2.2. ROTOROD SAMPLERROTOROD SAMPLER
3.3. BURKARD SAMPLERBURKARD SAMPLER
4.4. MEDIA (Nutrient agar, Czapek’s Dox Agar)MEDIA (Nutrient agar, Czapek’s Dox Agar)
5.5. ANDERSEN SAMPLERANDERSEN SAMPLER
6.6. CASCADE SAMPLERCASCADE SAMPLER
7.7. GAS CHROMATOGRAPHY- MASS SPECTROMETRYGAS CHROMATOGRAPHY- MASS SPECTROMETRY
FOR ENDOTOXINSFOR ENDOTOXINS
8.8. ELISA monoclonal antibodies for specific allergensELISA monoclonal antibodies for specific allergens
9.9. ERGOSTEROL/PEPTIDOGLYCAN QUANTIFICATIONERGOSTEROL/PEPTIDOGLYCAN QUANTIFICATION
BY IMMUNOLOGICAL TECHNIQUESBY IMMUNOLOGICAL TECHNIQUES
10.10. LIMULUS AMOEBOCYTE LYSATE (LAL) TEST FORLIMULUS AMOEBOCYTE LYSATE (LAL) TEST FOR
ENDOTOXINSENDOTOXINS

20. Handy sampler for dust

21. Centrifugal air sampler for bioaerosols

22. Colonies on agar strips

23. DETECTION OF ALLERGYDETECTION OF ALLERGY
►PRICK TESTPRICK TEST
►NASAL CHALLENGE TESTNASAL CHALLENGE TEST
►INTRADERMAL TEST >3 mm dia wheel isINTRADERMAL TEST >3 mm dia wheel is
positive.positive.
►ELISA / RAST TESTELISA / RAST TEST

24. SYMPTOMS OF ALLERGYSYMPTOMS OF ALLERGY
►RHITINITISRHITINITIS
►URTICARIAURTICARIA
►SNEEZING, DRY COUGHSNEEZING, DRY COUGH
►CONJUNCTIVITISCONJUNCTIVITIS
►DIARRHOEA/CRAMPSDIARRHOEA/CRAMPS
►WATERY DISCHARGEWATERY DISCHARGE
►ASTHMA, ECZEMAASTHMA, ECZEMA
►GRANULOMATOSISGRANULOMATOSIS
►PULMONARY FIBROSISPULMONARY FIBROSIS

25. LOW MOLECULAR WEIGHTLOW MOLECULAR WEIGHT
INDUSTRIAL ALLERGENSINDUSTRIAL ALLERGENS
►HAPTENS- bind with human endogenusHAPTENS- bind with human endogenus
protein and become Neoantigens-protein and become Neoantigens-
immunologic potentialimmunologic potential
1. POLY ISOCYANATES1. POLY ISOCYANATES
2. ACID ANHYDRIDES2. ACID ANHYDRIDES
3. REACTIVE AZO DYES3. REACTIVE AZO DYES
4.SULPHONO CHLORIDE4.SULPHONO CHLORIDE
5.PLATINUM SALTS5.PLATINUM SALTS
6. BERYLLIUM6. BERYLLIUM

26. TREATMENTTREATMENT
►Avoidance of allergensAvoidance of allergens
►Antihistamine – loratidine, fexafenadineAntihistamine – loratidine, fexafenadine
►CorticosteroidsCorticosteroids
►BronchodilatersBronchodilaters
►TheophyllineTheophylline
►CromonesCromones
►Immunosuppresents – cyclosporineImmunosuppresents – cyclosporine
►Immunotherapy- allergen vaccinationImmunotherapy- allergen vaccination
►Peptide immunotherapyPeptide immunotherapy

27. PREVENTIONPREVENTION
► Reduction of Dust at workplace by good ventilation andReduction of Dust at workplace by good ventilation and
exhaust system and good manufacturing practice.exhaust system and good manufacturing practice.
► Storing of plant raw material at low temperature andStoring of plant raw material at low temperature and
humidity that prevent growth of microorganisms.humidity that prevent growth of microorganisms.
► Ionization of air, cleaner production room, working of rawIonization of air, cleaner production room, working of raw
materials or sterilization for inactivation of endotoxinmaterials or sterilization for inactivation of endotoxin
► Postive pressure helmets during workPostive pressure helmets during work
► Remote control production process in agricultural andRemote control production process in agricultural and
wood industrywood industry
► Health education in agricultural and industrial workersHealth education in agricultural and industrial workers
► Avoidance or strict barriers to avoid contactAvoidance or strict barriers to avoid contact
► Government intervention for good practices. Example,Government intervention for good practices. Example,
passive indoor tobacco smoke can interfere in air quality.passive indoor tobacco smoke can interfere in air quality.