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  1. Norbäck D, Hashim Z, Ali F, Hashim JH
    Environ Res, 2021 06;197:111061.
    PMID: 33785322 DOI: 10.1016/j.envres.2021.111061
    Little is known on respiratory effects of indoor chemicals in the tropics. We investigated associations between asthma and respiratory infections in Malaysian students and chemical exposure at home and at school. Moreover, we investigated differences in home environment between the three main ethnic groups in Malaysia (Malay, Chinese, Indian). Totally, 462 students from 8 junior high schools in Johor Bahru participated (96% participation rate). The students answered a questionnaire on health and home environment. Climate, carbon dioxide (CO2), volatile organic compounds (VOC), formaldehyde and nitrogen dioxide (NO2) were measured inside and outside the schools. Multilevel logistic regression was applied to study associations between exposure and health. Totally 4.8% were smokers, 10.3% had wheeze, 9.3% current asthma, and had 18.8% any respiratory infection in the past 3 months. Malay students had more dampness or mould (p 
  2. Norbäck D, Hashim JH, Hashim Z, Ali F
    Sci Total Environ, 2017 Aug 15;592:153-160.
    PMID: 28319702 DOI: 10.1016/j.scitotenv.2017.02.215
    This paper studied associations between volatile organic compounds (VOC), formaldehyde, nitrogen dioxide (NO2) and carbon dioxide (CO2) in schools in Malaysia and rhinitis, ocular, nasal and dermal symptoms, headache and fatigue among students. Pupils from eight randomly selected junior high schools in Johor Bahru, Malaysia (N=462), participated (96%). VOC, formaldehyde and NO2 were measured by diffusion sampling (one week) and VOC also by pumped air sampling during class. Associations were calculated by multi-level logistic regression adjusting for personal factors, the home environment and microbial compounds in the school dust. The prevalence of weekly rhinitis, ocular, throat and dermal symptoms were 18.8%, 11.6%, 15.6%, and 11.1%, respectively. Totally 20.6% had weekly headache and 22.1% fatigue. Indoor CO2 were low (range 380-690 ppm). Indoor median NO2 and formaldehyde concentrations over one week were 23μg/m3 and 2.0μg/m3, respectively. Median indoor concentration of toluene, ethylbenzene, xylene, and limonene over one week were 12.3, 1.6, 78.4 and 3.4μg/m3, respectively. For benzaldehyde, the mean indoor concentration was 2.0μg/m3 (median<1μg/m3). Median indoor levels during class of benzene and cyclohexane were 4.6 and 3.7μg/m3, respectively. NO2 was associated with ocular symptoms (p<0.001) and fatigue (p=0.01). Formaldehyde was associated with ocular (p=0.004), throat symptoms (p=0.006) and fatigue (p=0.001). Xylene was associated with fatigue (p<0.001) and benzaldehyde was associated with headache (p=0.03). In conclusion, xylene, benzaldehyde, formaldehyde and NO2 in schools can be risk factors for ocular and throat symptoms and fatigue among students in Malaysia. The indoor and outdoor levels of benzene were often higher than the EU standard of 5μg/m3.
  3. Norbäck D, Hashim JH, Hashim Z, Wieslander G
    Int J Environ Health Res, 2024 Jan;34(1):213-224.
    PMID: 36335594 DOI: 10.1080/09603123.2022.2143482
    We studied associations between fractional exhaled nitric oxide (FeNO), health and household exposure among school children (N = 348) in Penang, Malaysia. Multiple logistic regression and linear mixed models were applied. Overall, 46.0% had elevated FeNO (>20 ppb) and 10.6% diagnosed asthma. Male gender (p = 0.002), parental asthma or allergy (p = 0.047), cat allergy (p = 0.009) and seafood allergy (p 
  4. Cai GH, Hashim JH, Hashim Z, Ali F, Bloom E, Larsson L, et al.
    Pediatr Allergy Immunol, 2011 May;22(3):290-7.
    PMID: 21457336 DOI: 10.1111/j.1399-3038.2010.01127.x
    While there is a large variation of prevalence of asthma symptoms worldwide, what we do know is that it is on the rise in developing countries. However, there are few studies on allergens, moulds and mycotoxin exposure in schools in tropical countries. The aims were to measure selected fungal DNA, furry pet allergens and mycotoxins in dust samples from schools in Malaysia and to study associations with pupils' respiratory health effects. Eight secondary schools and 32 classrooms in Johor Bahru, Malaysia were randomly selected. A questionnaire with standardized questions was used for health assessment in 15 randomly selected pupils from each class. The school buildings were inspected and both indoor and outdoor climate were measured. Dust samples were collected by cotton swabs and Petri dishes for fungal DNA, mycotoxins and allergens analysis. The participation rate was 96% (462/480 invited pupils), with a mean age of 14 yr (range 14-16). The pupils mostly reported daytime breathlessness (41%), parental asthma or allergy (22%), pollen or pet allergy (21%) and doctor-diagnosed asthma (13%) but rarely reported night-time breathlessness (7%), asthma in the last 12 months (3%), medication for asthma (4%) or smoking (5%). The inspection showed that no school had any mechanical ventilation system, but all classrooms had openable windows that were kept open during lectures. The mean building age was 16 yr (range 3-40) and the mean indoor and outdoor CO(2) levels were 492 ppm and 408 ppm, respectively. The mean values of indoor and outdoor temperature and relative humidity were the same, 29°C and 70% respectively. In cotton swab dust samples, the Geometric Mean (GM) value for total fungal DNA and Aspergillus/Penicillium (Asp/Pen) DNA in swab samples (Cell Equivalents (CE)/m(2)) was 5.7*10(8) and 0.5*10(8), respectively. The arithmetic mean (CE/m(2)) for Aspergillus versicolor DNA was 8780, Stachybotrys chartarum DNA was 26 and Streptomyces DNA was 893. The arithmetic means (pg/m(2)) for the mycotoxins sterigmatocystin and verrucarol were 2547 and 17, respectively. In Petri dish dust samples, the GM value for total fungal DNA and Asp/Pen DNA (CE/m(2) per day) was 9.2*10(6) and 1.6*10(6), respectively. The arithmetic mean (CE/m(2) per day) for A. versicolor DNA was 1478, S. chartarum DNA was 105 and Streptomyces DNA was 1271, respectively. The GM value for cat (Fel d1) allergen was 5.9 ng/m(2) per day. There were positive associations between A. versicolor DNA, wheeze and daytime breathlessness and between Streptomyces DNA and doctor-diagnosed asthma. However, the associations were inverse between S. chartarum DNA and daytime breathlessness and between verrucarol and daytime breathlessness. In conclusion, fungal DNA and cat allergen contamination were common in schools from Malaysia and there was a high prevalence of respiratory symptoms among pupils. Moreover, there were associations between levels of some fungal DNA and reported respiratory health in the pupils.
  5. Lim FL, Hashim Z, Md Said S, Than LT, Hashim JH, Norbäck D
    J Asthma, 2016 Mar;53(2):170-8.
    PMID: 26300213 DOI: 10.3109/02770903.2015.1077861
    OBJECTIVE: There are few studies on fractional exhaled nitric oxide (FeNO) and respiratory symptoms among adults in tropical areas. The aim was to study associations between FeNO and selected personal factors, respiratory symptoms, allergies, office characteristics and indoor office exposures among office workers (n = 460) from a university in Malaysia.
    METHODS: Information on health was collected by a questionnaire, skin prick test and FeNO measurement. Temperature, relative air humidity, carbon monoxide and carbon dioxide were measured in the offices. Settled dust was vacuumed in the offices and analyzed for endotoxin, (1,3)-β-glucan and house dust mites allergens, namely Dermatophagoides pteronyssinus (Der p 1) and Dermatophagoides farinae (Der f 1). Two-level linear mixed models and multiple logistic regression were used to analyze the associations.
    RESULTS: One-fourth (25.9%) of the office workers had elevated FeNO level (≥ 25 ppb) and 61.5% had HDM, cat, seafood or pollen allergy. Male gender (p < 0.001), current smoking (p = 0.037), height (p < 0.001) and atopy (p < 0.001) were associated with FeNO. The amount of vacuumed dust was associated with FeNO among atopic subjects (p = 0.009). Asthma and rhinitis symptoms were associated with FeNO (p < 0.05), especially among atopic subjects. In particular, a combination of atopy and elevated FeNO were associated with doctor-diagnosed asthma (p < 0.001), rhinitis (p < 0.001) and airway symptoms last 12 months (p < 0.001).
    CONCLUSION: Gender, smoking, height and atopy are important risk factors for elevated FeNO levels. A combination of allergy testing and FeNO measurement could be useful in respiratory illness epidemiology studies and patient investigations in tropical areas.
    KEYWORDS: Adults; Malaysia; allergy; office; respiratory symptoms; rhinitis; tropical areas
  6. Norbäck D, Hashim JH, Markowicz P, Cai GH, Hashim Z, Ali F, et al.
    Sci Total Environ, 2016 Mar 1;545-546:95-103.
    PMID: 26745297 DOI: 10.1016/j.scitotenv.2015.12.072
    This paper studied associations between ocular symptoms, rhinitis, throat and dermal symptoms, headache and fatigue in students by ethnicity and in relation to exposure to chemical microbial markers and fungal DNA in vacuumed dust in schools in Malaysia. A total of 462 students from 8 randomly selected secondary schools in Johor Bahru, Malaysia, participated (96% response rate). Dust was vacuumed from 32 classrooms and analysed for levels of five types of endotoxin as 3-hydroxy fatty acids (C10, C12, C14, C16 and C18 3-OH), muramic acid, ergosterol and five sequences of fungal DNA. Multiple logistic regression was applied. Totally 11.9% reported weekly ocular symptoms, 18.8% rhinitis, 15.6% throat and 11.1% dermal symptoms, 20.6% headache and 22.1% tiredness. Totally 21.1% reported pollen or furry pet allergy (atopy) and 22.0% parental asthma or allergy. Chinese students had less headache than Malay and Indian had less rhinitis and less tiredness than Malay. Parental asthma/allergy was a risk factor for ocular (odds ratio=3.79) and rhinitis symptoms (OR=3.48). Atopy was a risk factor for throat symptoms (OR=2.66), headache (OR=2.13) and tiredness (OR=2.02). There were positive associations between amount of fine dust in the dust samples and ocular symptoms (p<0.001) and rhinitis (p=0.006). There were positive associations between C14 3-OH and rhinitis (p<0.001) and between C18 3-OH and dermal symptoms (p=0.007). There were negative (protective) associations between levels of total endotoxin (LPS) (p=0.004) and levels of ergosterol (p=0.03) and rhinitis and between C12 3-OH and throat symptoms (p=0.004). In conclusion, the amount of fine dust in the classroom was associated with rhinitis and other SBS symptoms and improved cleaning of the schools is important. Endotoxin in the school dust seems to be mainly protective for rhinitis and throat symptoms but different types of endotoxin could have different effects. The ethnic differences in symptoms among the students deserve further attention.
  7. Norbäck D, Hashim JH, Cai GH, Hashim Z, Ali F, Bloom E, et al.
    PLoS One, 2016;11(2):e0147996.
    PMID: 26829324 DOI: 10.1371/journal.pone.0147996
    There are few studies on rhinitis and sick building syndrome (SBS) among students in tropical countries. We studied associations between levels of five fungal DNA sequences, two mycotoxins (sterigmatocystin and verrucarol) and cat allergen (Fel d 1) levels in schools and rhinitis and other weekly SBS symptoms in the students. Fungal DNA was measured by quantitative PCR and cat allergen by ELISA. Pupils (N = 462) from eight randomly selected schools in Johor Bahru, Malaysia participated (96%). Dust samples were collected by cotton swabs and Petri dishes exposed for one week. None of the schools had a mechanical ventilation system, but all classrooms had openable windows that were kept open during lectures and indoor CO2 levels were low (mean 492 ppm; range 380-690 ppm). Weekly nasal symptoms (rhinitis) (18.8%), ocular (11.6%), throat (11.1%), dermal symptoms, headache (20.6%) and tiredness (22.1%) were common. Total fungal DNA in swab samples was associated with rhinitis (p = 0.02), ocular symptoms (p = 0.009) and tiredness (p = 0.001). There were positive associations between Aspergillus versicolor DNA in Petri dish samples, ocular symptoms (p = 0.02) and tiredness (p = 0.001). The level of the mycotoxin verrucarol (produced by Stachybotrys chartarum) in swab samples was positively associated with tiredness (p = 0.04). Streptomyces DNA in swab samples (p = 0.03) and Petri dish samples (p = 0.03) were negatively associated with tiredness. In conclusion, total fungal contamination, measured as total fungal DNA) in the classrooms, Aspergillus versicolor and verrucarol can be risk factors for rhinitis and SBS symptoms among students in the tropical country Malaysia.
  8. Lim FL, Hashim Z, Md Said S, Than LT, Hashim JH, Norbäck D
    Sci Total Environ, 2015 Dec 1;536:353-61.
    PMID: 26225741 DOI: 10.1016/j.scitotenv.2015.06.137
    There are few studies on sick building syndrome (SBS) including clinical measurements for atopy and fractional exhaled nitric oxide (FeNO). Our aim was to study associations between SBS symptoms, selected personal factors, office characteristics and indoor office exposures among office workers from a university in Malaysia. Health data were collected by a questionnaire (n=695), skin prick test (SPT) (n=463) and FeNO test (n=460). Office settled dust was vacuumed and analyzed for endotoxin, (1,3)-β-glucan and house dust mites (HDM) allergens group 1 namely Dermatophagoides pteronyssinus (Der p 1) and Dermatophagoides farinae (Der f 1). Office indoor temperature, relative air humidity (RH), carbon monoxide (CO) and carbon dioxide (CO2) were measured by a direct reading instrument. Associations were studied by two-levels multiple logistic regression with mutual adjustment and stratified analysis. The prevalence of weekly dermal, mucosal and general symptoms was 11.9%, 16.0% and 23.0% respectively. A combination of SPT positivity (allergy to HDM or cat) and high FeNO level (≥25 ppb) was associated with dermal (p=0.002), mucosal (p<0.001) and general symptoms (p=0.05). Der f1 level in dust was associated with dermal (p<0.001), mucosal (p<0.001) and general (p=0.02) symptoms. Among those with allergy to D. farinae, associations were found between Der f 1 levels in dust and dermal (p=0.003), mucosal (p=0.001) and general symptoms (p=0.007). Office-related symptoms were associated with Der f 1 levels in dust (p=0.02), low relative air humidity (p=0.04) and high office temperature (p=0.05). In conclusion, a combination of allergy to cat or HDM and high FeNO is a risk factor for SBS symptoms. Der f 1 allergen in dust can be a risk factor for SBS in the office environment, particularly among those sensitized to Der f 1 allergen.
  9. Lim FL, Hashim Z, Than LT, Md Said S, Hisham Hashim J, Norbäck D
    PLoS One, 2015;10(4):e0124905.
    PMID: 25923543 DOI: 10.1371/journal.pone.0124905
    A prevalence study was conducted among office workers in Malaysia (N= 695). The aim of this study was to examine associations between asthma, airway symptoms, rhinitis and house dust mites (HDM) and cat allergy and HDM levels in office dust. Medical data was collected by a questionnaire. Skin prick tests were performed for HDM allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae) and cat allergen Felis domesticus. Indoor temperature and relative air humidity (RH) were measured in the offices and vacuumed dust samples were analyzed for HDM allergens. The prevalence of D. pteronyssinus, D. farinae and cat allergy were 50.3%, 49.0% and 25.5% respectively. Totally 9.6% had doctor-diagnosed asthma, 15.5% had current wheeze and 53.0% had current rhinitis. The Der p 1 (from D. pteronyssinus) and Der f 1 (from D. farinae) allergens levels in dust were 556 ng/g and 658 ng/g respectively. Statistical analysis was conducted by multilevel logistic regression, adjusting for age, gender, current smoking, HDM or cat allergy, home dampness and recent indoor painting at home. Office workers with HDM allergy had more wheeze (p= 0.035), any airway symptoms (p= 0.032), doctor-diagnosed asthma (p= 0.005), current asthma (p= 0.007), current rhinitis (p= 0.021) and rhinoconjuctivitis (p< 0.001). Cat allergy was associated with wheeze (p= 0.021), wheeze when not having a cold (p= 0.033), any airway symptoms (p= 0.034), doctor-diagnosed asthma (p= 0.010), current asthma (p= 0.020) and nasal allergy medication (p= 0.042). Der f 1 level in dust was associated with daytime breathlessness (p= 0.033) especially among those with HDM allergy. Der f 1 levels were correlated with indoor temperature (p< 0.001) and inversely correlated with RH (p< 0.001). In conclusion, HDM and cat allergies were common and independently associated with asthma, airway symptoms and rhinitis. Der f 1 allergen can be a risk factor for daytime breathlessness.
  10. Ma'pol A, Hashim JH, Norbäck D, Weislander G, Hashim Z, Isa ZM
    J Asthma, 2020 08;57(8):842-849.
    PMID: 31155989 DOI: 10.1080/02770903.2019.1614614
    Background: Almost one third of the world population suffers from allergic conditions. Respiratory symptoms are common in Malaysian children but there are few studies on fractional exhaled nitric oxide (FeNO), inclusive of field clinical test for asthma among children in Malaysia. The aim was to provide insight on factors related to level of FeNO among students in Terengganu, Malaysia.Methods: In total, 487 randomly selected students from eight secondary schools participated (13-14 years old). A Standardized questionnaire was used to obtained information on doctors' diagnosed asthma, current asthma and respiratory symptoms. FeNO measurement and skin prick test (SPT to common allergen) were conducted.Results: The geometric mean FeNO was 16.7 ppb. Totally, 38.4% of students had elevated FeNO level (>20 ppb) and 40.3% had had positive SPT to house dust mites allergens (HDM), Dermatophagoides pteronyssinus (Der p 1), Dermatophagoides farinae (Der f 1) or Felis domisticus (cat). Male gender, height, parental history of allergy, self-reported allergy, and atopy were associated with FeNO. In particular, a combination of sensitization to HDM or cat and elevated FeNO were associated with doctor-diagnosed asthma and self-reported allergy to food, pollen and cat.Conclusion: Asthma, respiratory symptoms and sensitization to HDM and cat are common among students and presence of elevated FeNO levels indicate ongoing airway inflammation.
  11. Norbäck D, Hashim JH, Hashim Z, Cai GH, Sooria V, Ismail SA, et al.
    Sci Total Environ, 2017 Jan 15;577:148-154.
    PMID: 27802882 DOI: 10.1016/j.scitotenv.2016.10.148
    Few health studies exist on dampness and mould in schools in the tropics. We studied associations between fraction of exhaled nitric oxide (FeNO), respiratory symptoms and airway infections among students and dampness and fungal DNA in schools in Malaysia. A total of 368 randomly selected students from 32 classrooms in 8 secondary schools in Penang, Malaysia, participated (58% participation rate). Information on current respiratory symptoms and the home environment was collected by a standardised questionnaire. FeNO was measured by NIOX MINO (50ml/min). The classrooms were inspected and dust was collected by vacuuming on special filters and was analysed for five fungal DNA sequences by quantitative PCR. Linear mixed models and 3-level multiple logistic regression (school, classroom, student) were applied adjusting for demographic data and the home environment. Totally 10.3% reported doctor's diagnosed asthma, 15.1% current wheeze, 12.4% current asthma, 37.3% daytime breathlessness, 10.2% nocturnal breathlessness, 38.9% airway infections and 15.5% had pollen or furry pet allergy. The geometric mean of FeNO was 19.9ppb and 45% had elevated FeNO (>20ppb). Boys had higher levels of FeNO. Chinese had less daytime breathlessness than Malay (OR=0.30: p<0.001). Indoor carbon dioxide levels were low (380-720ppm). Dampness was observed in 18% of the classrooms and was associated with respiratory infections (OR=3.70; 95% CI 1.14-12.1) and FeNO (p=0.04). Aspergillus versicolor DNA was detected in 67% of the classrooms. Higher numbers of Aspergillus versicolor DNA in classroom dust were associated with wheeze (p=0.006), current asthma (p=0.002), respiratory infections (p=0.005) and elevated FeNO levels (p=0.02). In conclusion, respiratory symptoms were common among the students and the high FeNO levels indicate ongoing airway inflammation. Building dampness and the mould Aspergillus versicolor in schools in Malaysia can be risk factors for impaired respiratory health among the students.
  12. Lim FL, Hashim Z, Than LTL, Md Said S, Hashim JH, Norbäck D
    Int J Tuberc Lung Dis, 2019 11 01;23(11):1171-1177.
    PMID: 31718753 DOI: 10.5588/ijtld.18.0668
    OBJECTIVE: To examine the associations between endotoxin and (1,3)-β-glucan concentrations in office dust and respiratory symptoms and airway inflammation among 695 office workers in Malaysia.METHODS: Health data were collected using a questionnaire, sensitisation testing and measurement of fractional exhaled nitric oxide (FeNO). Indoor temperature, relative air humidity (RH) and carbon dioxide (CO₂) were measured in the offices and settled dust was vacuumed and analysed for endotoxin and (1,3)-β-glucan concentrations. Associations were analysed by two level multiple logistic regression.RESULTS: Overall, 9.6% of the workers had doctor-diagnosed asthma, 15.5% had wheeze, 18.4% had daytime attacks of breathlessness and 25.8% had elevated FeNO (≥25 ppb). The median levels in office dust were 11.3 EU/mg endotoxin and 62.9 ng/g (1,3)-β-glucan. After adjusting for personal and home environment factors, endotoxin concentration in dust was associated with wheeze (P = 0.02) and rhinoconjunctivitis (P = 0.007). The amount of surface dust (P = 0.04) and (1,3)-β-glucan concentration dust (P = 0.03) were associated with elevated FeNO.CONCLUSION: Endotoxin in office dust could be a risk factor for wheeze and rhinoconjunctivitis among office workers in mechanically ventilated offices in a tropical country. The amount of dust and (1,3)-β-glucan (a marker of indoor mould exposure) were associated with Th2 driven airway inflammation.
  13. Norbäck D, Hashim JH, Hashim Z, Sooria V, Ismail SA, Wieslander G
    Int J Hyg Environ Health, 2017 06;220(4):697-703.
    PMID: 28254266 DOI: 10.1016/j.ijheh.2017.01.016
    BACKGROUND: There are few studies on ocular effects of indoor mould exposure in schools, especially in the tropics OBJECTIVE: To study associations between eye symptoms and tear film break up time (BUT) in students and demographic data and fungal DNA in schools.

    METHODS: A school environment study was performed among randomly selected students in eight randomly selected secondary schools in Penang, Malaysia. Information on eye symptoms and demographic data was collected by a standardised questionnaire. BUT was measured by two methods, self-reported BUT (SBUT) and by the non-invasive Tearscope (NIBUT). Dust was collected by vacuuming in 32 classrooms and analysed for five fungal DNA sequences. Geometric mean (GM) for total fungal DNA was 7.31*104 target copies per gram dust and for Aspergillus/Penicillium DNA 3.34*104 target copies per gram dust. Linear mixed models and 3-level multiple logistic regression were applied adjusting for demographic factors.

    RESULTS: A total of 368 students (58%) participated and 17.4% reported weekly eye symptoms the last 3 months. The median SBUT and TBUT were 15 and 12s, respectively. Students wearing glasses (OR 2.41, p=0.01) and with a history of atopy (OR=2.67; p=0.008) had more eye symptoms. Girls had less eye symptoms than boys (OR=0.34; p=0.006) Indoor carbon dioxide in the classrooms was low (range 380-720ppm), temperature was 25-30°C and relative air humidity 70-88%. Total fungal DNA in vacuumed dust was associated with shorter SBUT (4s shorter per 105 target copies per gram dust; p=0.04) and NIBUT (4s shorter per 105 target copies per gram dust; p<0.001). Aspergillus/Penicillium DNA was associated with shorter NIBUT (5s shorter per 105 target copies per gram dust; p=0.01).

    CONCLUSION: Fungal contamination in schools in a tropical country can be a risk factor for impaired tear film stability among students.

  14. Norbäck D, Markowicz P, Cai GH, Hashim Z, Ali F, Zheng YW, et al.
    PLoS One, 2014;9(2):e88303.
    PMID: 24523884 DOI: 10.1371/journal.pone.0088303
    There are few studies on associations between respiratory health and allergens, fungal and bacterial compounds in schools in tropical countries. The aim was to study associations between respiratory symptoms in pupils and ethnicity, chemical microbial markers, allergens and fungal DNA in settled dust in schools in Malaysia. Totally 462 pupils (96%) from 8 randomly selected secondary schools in Johor Bahru, Malaysia, participated. Dust was vacuumed from 32 classrooms and analysed for levels of different types of endotoxin as 3-hydroxy fatty acids (3-OH), muramic acid, ergosterol, allergens and five fungal DNA sequences. Multiple logistic regression was applied. Totally 13.1% pupils reported doctor's diagnosed asthma, 10.3% wheeze and 21.1% pollen or pet allergy. Indian and Chinese children had less atopy and asthma than Malay. Carbon dioxide levels were low (380-690 ppm). No cat (Fel d1), dog (Can f 1) or horse allergens (Ecu cx) were detected. The levels of Bloomia tropicalis (Blo t), house dust mite allergens (Der p 1, Der f 1, Der m 1) and cockroach allergens (Per a 1 and Bla g 1) were low. There were positive associations between levels of Aspergillus versicolor DNA and daytime breathlessness, between C14 3-OH and respiratory infections and between ergosterol and doctors diagnosed asthma. There were negative (protective) associations between levels of C10 3-OH and wheeze, between C16 3-OH and day time and night time breathlessness, between cockroach allergens and doctors diagnosed asthma. Moreover there were negative associations between amount of fine dust, total endotoxin (LPS) and respiratory infections. In conclusion, endotoxin at school seems to be mainly protective for respiratory illness but different types of endotoxin could have different effects. Fungal contamination measured as ergosterol and Aspergillus versicolor DNA can be risk factors for respiratory illness. The ethnical differences for atopy and asthma deserve further attention.
  15. Fu X, Norbäck D, Yuan Q, Li Y, Zhu X, Hashim JH, et al.
    Sci Total Environ, 2021 Jan 20;753:141904.
    PMID: 32890872 DOI: 10.1016/j.scitotenv.2020.141904
    Sick building syndrome (SBS) is a collection of nonspecific syndromes linked with the built environment. The occurrence of SBS is associated with humidity, ventilation, moulds and microbial compounds exposure. However, no study has reported the association between indoor microbiome and SBS. In this study, 308 students were surveyed for SBS symptoms from 21 classrooms of 7 junior high schools from Johor Bahru, Malaysia, and vacuum dust from floor, desks and chairs was collected. High throughput amplicon sequencing (16S rRNA gene and ITS region) and quantitative PCR were conducted to characterize the absolute concentration of bacteria and fungi taxa. In total, 326 bacterial and 255 fungal genera were detected in dust with large compositional variation among classrooms. Also, half of these samples showed low compositional similarity to microbiome data deposited in the public database. The number of observed OTUs in Gammaproteobacteria was positively associated with SBS (p = 0.004). Eight microbial genera were associated with SBS (p 
  16. Isa KNM, Jalaludin J, Elias SM, Than LTL, Jabbar MA, Saudi ASM, et al.
    Ecotoxicol Environ Saf, 2021 Sep 15;221:112430.
    PMID: 34147866 DOI: 10.1016/j.ecoenv.2021.112430
    The exposure of school children to indoor air pollutants has increased allergy and respiratory diseases. The objective of this study were to determine the toxicodynamic interaction of indoor pollutants exposure, biological and chemical with expression of adhesion molecules on eosinophil and neutrophil. A self-administered questionnaire, allergy skin test, and fractional exhaled nitric oxide (FeNO) analyser were used to collect information on health status, sensitization to allergens and respiratory inflammation, respectively among school children at age of 14 years. The sputum induced were analysed to determine the expression of CD11b, CD35, CD63 and CD66b on eosinophil and neutrophil by using flow cytometry technique. The particulate matter (PM2.5 and PM10), NO2, CO2, and formaldehyde, temperature, and relative humidity were measured inside the classrooms. The fungal DNA were extracted from settled dust collected from classrooms and evaluated using metagenomic techniques. We applied chemometric and regression in statistical analysis. A total of 1869 unique of operational taxonomic units (OTUs) of fungi were identified with dominated at genus level by Aspergillus (15.8%), Verrucoconiothyrium (5.5%), and Ganoderma (4.6%). Chemometric and regression results revealed that relative abundance of T. asahii were associated with down regulation of CD66b expressed on eosinophil, and elevation of FeNO levels in predicting asthmatic children with model accuracy of 63.6%. Meanwhile, upregulation of CD11b expressed on eosinophil were associated with relative abundance of A. clavatus and regulated by PM2.5. There were significant association of P. bandonii with upregulation of CD63 expressed on neutrophil and exposure to NO2. Our findings indicate that exposure to PM2.5, NO2, T. asahii, P.bandonii and A.clavatus are likely interrelated with upregulation of activation and degranulation markers on both eosinophil and neutrophil.
  17. Sun Y, Zhang M, Ou Z, Meng Y, Chen Y, Lin R, et al.
    Eur Respir J, 2022 Nov;60(5).
    PMID: 35618276 DOI: 10.1183/13993003.00260-2022
    BACKGROUND: Indoor microbial exposure is associated with asthma, but the health effects of indoor metabolites and chemicals have not been comprehensively assessed.

    METHODS: We collected classroom dust from 24 junior high schools in three geographically distanced areas in Malaysia (Johor Bahru, Terengganu and Penang), and conducted culture-independent high-throughput microbiome and untargeted metabolomics/chemical profiling.

    RESULTS: 1290 students were surveyed for asthma symptoms (wheeze). In each centre, we found significant variation in the prevalence of wheeze among schools, which could be explained by personal characteristics and air pollutants. Large-scale microbial variations were observed between the three centres; the potential protective bacteria were mainly from phyla Actinobacteria in Johor Bahru, Cyanobacteria in Terengganu and Proteobacteria in Penang. In total, 2633 metabolites and chemicals were characterised. Many metabolites were enriched in low-wheeze schools, including plant secondary metabolites flavonoids/isoflavonoids (isoliquiritigenin, formononetin, astragalin), indole and derivatives (indole, serotonin, 1H-indole-3-carboxaldehyde), and others (biotin, chavicol). A neural network analysis showed that the indole derivatives were co-occurring with the potential protective microbial taxa, including Actinomycetospora, Fischerella and Truepera, suggesting these microorganisms may pose health effects by releasing indole metabolites. A few synthetic chemicals were enriched in high-wheeze schools, including pesticides (2(3H)-benzothiazolethione), fragrances (2-aminobenzoic acid, isovaleric acid), detergents and plastics (phthalic acid), and industrial materials (4,4-sulfonyldiphenol).

    CONCLUSIONS: This is the first association study between high-throughput indoor chemical profiling and asthma symptoms. The consistent results from the three centres indicate that indoor metabolites/chemicals could be a better indicator than the indoor microbiome for environmental and health assessments, providing new insights for asthma prediction, prevention and control.

  18. Fu X, Norbäck D, Yuan Q, Li Y, Zhu X, Hashim JH, et al.
    Environ Int, 2020 05;138:105664.
    PMID: 32200316 DOI: 10.1016/j.envint.2020.105664
    Indoor microbial diversity and composition are suggested to affect the prevalence and severity of asthma by previous home microbiome studies, but no microbiome-health association study has been conducted in a school environment, especially in tropical countries. In this study, we collected floor dust and environmental characteristics from 21 classrooms, and health data related to asthma symptoms from 309 students, in junior high schools in Johor Bahru, Malaysia. The bacterial and fungal composition was characterized by sequencing 16s rRNA gene and internal transcribed spacer (ITS) region, and the absolute microbial concentration was quantified by qPCR. In total, 326 bacterial and 255 fungal genera were characterized. Five bacterial (Sphingobium, Rhodomicrobium, Shimwellia, Solirubrobacter, Pleurocapsa) and two fungal (Torulaspora and Leptosphaeriaceae) taxa were protective for asthma severity. Two bacterial taxa, Izhakiella and Robinsoniella, were positively associated with asthma severity. Several protective bacterial taxa including Rhodomicrobium, Shimwellia and Sphingobium have been reported as protective microbes in previous studies, whereas other taxa were first time reported. Environmental characteristics, such as age of building, size of textile curtain per room volume, occurrence of cockroaches, concentration of house dust mite allergens transferred from homes by the occupants, were involved in shaping the overall microbial community but not asthma-associated taxa; whereas visible dampness and mold, which did not change the overall microbial community for floor dust, was negatively associated with the concentration of protective bacteria Rhodomicrobium (β = -2.86, p = 0.021) of asthma. The result indicates complex interactions between microbes, environmental characteristics and asthma symptoms. Overall, this is the first indoor microbiome study to characterize the asthma-associated microbes and their environmental determinant in the tropical area, promoting the understanding of microbial exposure and respiratory health in this region.
  19. Fu X, Yuan Q, Zhu X, Li Y, Meng Y, Hashim JH, et al.
    Environ Sci Process Impacts, 2021 Aug 01;23(8):1171-1181.
    PMID: 34278392 DOI: 10.1039/d1em00115a
    Pathogens are commonly present in the human respiratory tract, but symptoms are varied among individuals. The interactions between pathogens, commensal microorganisms and host immune systems are important in shaping the susceptibility, development and severity of respiratory diseases. Compared to the extensive studies on the human microbiota, few studies reported the association between indoor microbiome exposure and respiratory infections. In this study, 308 students from 21 classrooms were randomly selected to survey the occurrence of respiratory infections in junior high schools of Johor Bahru, Malaysia. Vacuum dust was collected from the floor, chairs and desks of these classrooms, and high-throughput amplicon sequencing (16S rRNA and ITS) and quantitative PCR were conducted to characterize the absolute concentration of the indoor microorganisms. Fifteen bacterial genera in the classes Actinobacteria, Alphaproteobacteria, and Cyanobacteria were protectively associated with respiratory infections (p < 0.01), and these bacteria were mainly derived from the outdoor environment. Previous studies also reported that outdoor environmental bacteria were protectively associated with chronic respiratory diseases, such as asthma, but the genera identified were different between acute and chronic respiratory diseases. Four fungal genera from Ascomycota, including Devriesia, Endocarpon, Sarcinomyces and an unclassified genus from Herpotrichillaceae, were protectively associated with respiratory infections (p < 0.01). House dust mite (HDM) allergens and outdoor NO2 concentration were associated with respiratory infections and infection-related microorganisms. A causal mediation analysis revealed that the health effects of HDM and NO2 were partially or fully mediated by the indoor microorganisms. This is the first study to explore the association between environmental characteristics, microbiome exposure and respiratory infections in a public indoor environment, expanding our understanding of the complex interactions among these factors.
  20. Fu X, Du B, Meng Y, Li Y, Zhu X, Ou Z, et al.
    PMID: 36883483 DOI: 10.1039/d2em00480a
    Rhinitis is one of the most prevalent chronic diseases globally. Microbiome exposure affects the occurrence of rhinitis. However, previous studies did not differentiate allergic rhinitis (AR) and non-allergic rhinitis (NAR) in the microbial association analysis. In this study, we investigate 347 students in 8 junior high schools, Terengganu, Malaysia, who were categorized as healthy (70.9%), AR (13.8%) and NAR (15.3%) based on a self-administered questionnaire and skin prick tests of pollen, pet, mould and house dust mite allergens. Classroom microbial and metabolite exposure in vacuumed dust was characterized by PacBio long-read amplicon sequencing, quantitative PCR and LC-MS-based untargeted metabolomics. Our findings indicate a similar microbial association pattern between AR and NAR. The richness in Gammaproteobacteria was negatively associated with AR and NAR symptoms, whereas total fungal richness was positively associated with AR and NAR symptoms (p < 0.05). Brasilonema bromeliae and Aeromonas enteropelogenes were negatively associated with AR and NAR, and Deinococcus was positively associated with AR and NAR (p < 0.01). Pipecolic acid was protectively associated with AR and NAR symptoms (OR = 0.06 and 0.13, p = 0.009 and 0.045). A neural network analysis showed that B. bromeliae was co-occurring with pipecolic acid, suggesting that the protective role of this species may be mediated by releasing pipecolic acid. Indoor relative humidity and the weight of vacuum dust were associated with AR and NAR, respectively (p < 0.05), but the health effects were mediated by two protective bacterial species, Aliinostoc morphoplasticum and Ilumatobacter fluminis. Overall, our study reported a similar microbial association pattern between AR and NAR and also revealed the complex interactions between microbial species, environmental characteristics, and rhinitis symptoms.
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