Microbial growth in building material samples and occupants' health in severely moisture-damaged homes.

There is no commonly approved approach to detect and quantify the health-relevant microbial exposure in moisture-damaged buildings. In 39 single-family homes with severe moisture damage, we studied whether concentrations of viable microbes in building material samples are associated with health among 71 adults and 68 children, and assessed with symptoms questionnaires, exhaled NO, and peak expiratory flow (PEF) variability. Symptoms were grouped into three scores: upper respiratory symptoms, lower respiratory symptoms, and general symptoms. The homes were divided into three groups based on viable counts of fungi, actinomycetes, and total bacteria cultivated from building material samples. Highest group of actinomycete counts was associated with more general symptoms, worse perceived health, and higher daily PEF variability (aOR 12.51; 1.10-141.90 as compared to the lowest group) among adults, and with an increase in lower respiratory symptoms in children, but the confidence intervals were wide. We observed significant associations of fungal counts and total microbial score with worse perceived health in adults. No associations with exhaled NO were observed.

Indoor fungi: companions and contaminants.

This review discusses the role of fungi and fungal products in indoor environments, especially as agents of human exposure. Fungi are present everywhere, and knowledge for indoor environments is extensive on their occurrence and ecology, concentrations, and determinants. Problems of dampness and mold have dominated the discussion on indoor fungi. However, the role of fungi in human health is still not well understood. In this review, we take a look back to integrate what cultivation-based research has taught us alongside more recent work with cultivation-independent techniques. We attempt to summarize what is known today and to point out where more data is needed for risk assessment associated with indoor fungal exposures. New data have demonstrated qualitative and quantitative richness of fungal material inside and outside buildings. Research on mycotoxins shows that just as microbes are everywhere in our indoor environments, so too are their metabolic products. Assessment of fungal exposures is notoriously challenging due to the numerous factors that contribute to the variation of fungal concentrations in indoor environments. We also may have to acknowledge and incorporate into our understanding the complexity of interactions between multiple biological agents in assessing their effects on human health and well-being.

Determinants, reproducibility, and seasonal variation of bacterial cell wall components and viable counts in house dust.

The objectives of this study were (i) to assess the determinants that affect concentrations of the bacterial cell wall components 3-hydroxy fatty acids (3-OH FAs) and muramic acid and of total viable bacteria and actinomycetes in house dust; and (ii) to examine the seasonal variation and reproducibility of these bacterial cell wall components in house dust. A number of lifestyle and environmental factors, mostly not consistent for different bacterial measures but commonly including the type of dwelling and farming (number of livestock), explained up to 37% of the variation of the bacterial concentrations in 212 homes in Eastern Finland. The reproducibility of 3-OH FAs and muramic acid measurements in house dust were studied in five urban homes and were found to be generally high (ICC 74-84%). Temporal variation observed in repeated sampling of the same home throughout a year was more pronounced for 3-OH FAs determinations (ICC 22%) than for muramic acid (ICC 55-66%). We conclude that determinants vary largely for different types of bacterial measurements in house dust; the measured parameters represent different aspects of the bacterial content indoors. More than one sample is needed to describe bacterial concentrations in house dust in the home environment due to large temporal variation.

Determinants, reproducibility, and seasonal variation of ergosterol levels in house dust.

UNLABELLED: This study aimed to clarify the determinants that affect the concentrations of ergosterol and viable fungi in house dust and to examine the seasonal variation and reproducibility of ergosterol concentrations indoors. In studying the determinants, dust samples from living room floors and vacuum cleaner dust bags were collected from 107 farming and 105 non-farming homes. Ergosterol levels were determined with gas chromatography-mass spectrometry,and the dust bag dust was cultivated for enumeration of fungal genera. Lifestyle and environmental factors, for example using of the fireplace, and visible mold observations in homes, explained 20­26% of the variation of fungal concentrations. For the reproducibility study, samples were collected from five urban homes in four different seasons. The reproducibility of ergosterol determinations within a sample was excellent (ICC = 89.8) for floor dust and moderate (ICC = 63.8) for dust bag dust, but poor when sampling the same home throughout a year (ICC = 31.3 and 12.6, respectively) due to large temporal variation in ergosterol concentrations. In conclusion, environmental characteristics only partially predicted the variation of fungal concentrations. Based on these studies, we recommend repeated sampling of dust over time if one seeks to adequately describe overall fungal levels and exposure in a home. PRACTICAL IMPLICATIONS: This study shows that levels of ergosterol and viable fungi in house dust are related to visible mold observations. Only 20% of the variation in fungal levels can be explained with questionnaires, and therefore, environmental samples need to be taken in addition. Reproducibility of ergosterol determination was excellent for floor dust, and thus, ergosterol measurements from floor dust samples could be suitable for assessing the fungal load in building investigations. The temporal variation needs to be taken into account when describing the ergosterol concentration of urban homes.

Respiratory health and endotoxin: associations and modification by CD14/-260 genotype.

Exposure to endotoxin has been associated with increased respiratory symptoms and decrements in lung function in occupational settings but little is known about the health effects of domestic exposure in adults. Here, we describe the association of respiratory disease, immunoglobulin (Ig)E sensitisation, bronchial reactivity and lung function with mattress endotoxin levels in adults, and determine whether these associations are modified by polymorphisms in CD14. Endotoxin levels in mattress dust from a population-based sample of 972 adults were measured. Associations were examined using generalised linear mixed models, adjusting for individual and household confounders. Effect modification of these associations by CD14/-260 (rs2569190) was assessed. Mattress endotoxin levels varied from 0.1 to 402.6 EU · mg(-1). Although there was no overall association of lung function with endotoxin exposure, there was evidence that the association of forced expiratory volume in 1 s and forced vital capacity with endotoxin was modified by CD14/-260 genotype (p-value for interaction 0.005 and 0.013, respectively). There was no evidence that symptoms, IgE sensitisation or bronchial reactivity were associated with mattress endotoxin levels. In this large epidemiological study of adults, there was no evidence that mattress endotoxin level was associated with respiratory symptoms or IgE sensitisation but the association of lung function with endotoxin levels may be modified by CD14 genotype.

Occurrence of moisture problems in schools in three countries from different climatic regions of Europe based on questionnaires and building inspections - the HITEA study.

UNLABELLED: The aim of this study was to assess occurrence of dampness and mold in school buildings in three European countries (the Netherlands, Spain, and Finland), representing different climatic regions. An assessment was performed utilizing both questionnaires and on-site building investigations, and the agreement between these two methods was evaluated for validation purposes. On the basis of questionnaire data from a representative sample of schools, different types of moisture problems were reported in 24-47% of all school buildings at the time of the study. Most commonly reported was dampness in the Netherlands, moisture/water damage in Spain, and mold odor in Finland. Subsequently, 20-24 schools per country were selected for on-site inspections by trained staff. The overall agreement between the questionnaire and inspection data was good (kappa-value 0.62), however, with large differences (0.39-0.91) between countries. Extrapolating from the inspection data, the minimum estimates for prevalence of moisture problems in school buildings are 20% in the Netherlands, 41% in Spain, and 24% in Finland. In conclusion, moisture problems (such as moisture damage, dampness, and mold) are relatively common in schools. The occurrence and severity may vary across geographical areas, which can be partly explained by building characteristics. PRACTICAL IMPLICATIONS: On the basis of this study, the prevalence of verified moisture problems in school buildings was highest in Spain, but lower and similar in Finland and the Netherlands. Questionnaire-based surveys can be used to assess moisture problems in school buildings, but because of large variation in agreement with inspection data, the questionnaire needs to be validated by on-site inspections in a subsample of the surveyed buildings.

Co-occurrence of toxic bacterial and fungal secondary metabolites in moisture-damaged indoor environments.

UNLABELLED: Toxic microbial secondary metabolites have been proposed to be related to adverse health effects observed in moisture-damaged buildings. Initial steps in assessing the actual risk include the characterization of the exposure. In our study, we applied a multi-analyte tandem mass spectrometry-based methodology on sample materials of severely moisture-damaged homes, aiming to qualitatively and quantitatively describe the variety of microbial metabolites occurring in building materials and different dust sample types. From 69 indoor samples, all were positive for at least one of the 186 analytes targeted and as many as 33 different microbial metabolites were found. For the first time, the presence of toxic bacterial metabolites and their co-occurrence with mycotoxins were shown for indoor samples. The bacterial compounds monactin, nonactin, staurosporin and valinomycin were exclusively detected in building materials from moist structures, while chloramphenicol was particularly prevalent in house dusts, including settled airborne dust. These bacterial metabolites are highly bioactive compounds produced by Streptomyces spp., a group of microbes that is considered a moisture damage indicator in indoor environments. We show that toxic bacterial metabolites need to be considered as being part of very complex and diverse microbial exposures in 'moldy' buildings. PRACTICAL IMPLICATIONS: Bacterial toxins co-occur with mycotoxins in moisture-damaged indoor environments. These compounds are measurable also in settled airborne dust, indicating that inhalation exposure takes place. In attempts to characterize exposures to microbial metabolites not only mycotoxins but also bacterial metabolites have to be targeted by the analytical methods applied. We recommend including analysis of samples of outdoor air in the course of future indoor assessments, in an effort to better understand the outdoor contribution to the indoor presence of microbial toxins. There is a need for a sound risk assessment concerning the exposure to indoor microbial toxins at concentrations detectable in moisture-damaged indoor environments.

Survival of amoebae on building materials.

UNLABELLED: Moisture damage and concurrent microbial growth in buildings are associated with adverse health effects among the occupants. However, the causal agents for the symptoms are unclear although microbes are assumed to play a major role. Fungi and bacteria are not the only microbes inhabiting moist building materials; it was recently revealed that amoebae are also present. As amoebae have the potential to harbor many pathogens and to modulate the characteristics of growing microbes, a better appreciation of the growth and survival of amoebae in moisture damage conditions will add to the understanding of their effects on health outcomes. In this study, we investigated the ability of amoebae to survive on six building materials. Furthermore, both aged and unused materials were tested. Amoebae survived on gypsum board and mineral wool for the whole 2 months experiment even without additional sustenance. When sustenance (heat-killed bacteria) was available, aged pine wood and birch wood also allowed their survival. In contrast, amoebae were quickly killed on fresh pine wood and they did not survive on concrete or linoleum. In conclusion, our data show that amoebae can persist on several common building materials once these materials become wet. PRACTICAL IMPLICATIONS: Amoebae are able to survive on many building materials should the materials become wet. Amoebae have the potential to increase growth, cytotoxicity, and pathogenicity of other microbes present in moisture damages, and they may carry potentially pathogenic bacteria as endosymbionts and thus introduce them into the indoor air. Therefore, amoebae may have a prominent role in the microbial exposures occurring in moisture-damaged buildings. The presence of amoebae could be usefully included in reporting the microbial damage of material samples.

Exposure to environmental bacteria may have differing effects on tumour necrosis factor-alpha and interleukin-6-producing capacity in infancy.

BACKGROUND: Our previous study showed an association between increased concentration of endotoxin in house dust and elevated IFN-gamma responses in neonates. The impact of other microbial agents on immune responses in infancy is poorly known. OBJECTIVE: To examine whether stimulated cytokine responses of mothers and their children are associated with concentrations of other microbial markers in addition to endotoxin in house dust samples. METHODS: Mitogen-stimulated production of IFN-gamma, IL-4, IL-6 and TNF-alpha was measured in cord blood and in peripheral blood of mothers (n=29) and their children (n=29) 3 months after birth. Gas chromatography mass spectrometric analysis was applied to measure the concentrations of ergosterol (marker of fungal biomass), muramic acid (indicating the presence of Gram-positive bacteria) and 3-hydroxy fatty acids (C(10:0)-C(14:0), indicating the presence of Gram-negative bacteria) in house dust. Endotoxin was determined with Limulus assay. RESULTS: Significant mother-to-child correlations were observed in stimulated production of TNF-alpha and IL-6 3 months after birth. 3-hydroxy fatty acid (C(10:0)-C(14:0)) levels in bed dust were inversely associated with the production of TNF-alpha and IL-6 in blood samples of mothers and their 3-month-old children. High concentrations of muramic acid in floor dust were related to increased production of TNF-alpha and IL-6 at the age of 3 months. In contrast to endotoxin, none of the other microbial markers were significantly associated with enhanced IFN-gamma-producing capacity from birth to 3 months. CONCLUSIONS: Exposure to Gram-negative bacteria and their components may be associated with down-regulated immune responses in early infancy, indicated as an impaired production of pro-inflammatory cytokines following mitogen stimulation. Gram-positive bacteria and their constituents seem to have opposite effects. Of the measured markers, exposure to bioactive endotoxin appears to have the strongest impact on T-helper type 1 responses.

Analysis of fungal flora in indoor dust by ribosomal DNA sequence analysis, quantitative PCR, and culture.

In recent years increasing attention has been given to the potential health effects of fungal exposure in indoor environments. We used large-scale sequencing of the fungal internal transcribed spacer region (ITS) of nuclear ribosomal DNA to describe the mycoflora of two office buildings over the four seasons. DNA sequencing was complemented by cultivation, ergosterol determination, and quantitative PCR analyses. Sequences of 1,339 clones were clustered into 394 nonredundant fungal operational taxonomical units containing sequences from 18 fungal subclasses. The observed flora differed markedly from that recovered by cultivation, the major differences being the near absence of several typical indoor mold genera such as Penicillium and Aspergillus spp. and a high prevalence of basidiomycetes in clone libraries. A total of 55% of the total diversity constituted of unidentifiable ITS sequences, some of which may represent novel fungal species. Dominant species were Cladosporium cladosporioides and C. herbarum, Cryptococcus victoriae, Leptosphaerulina americana and L. chartarum, Aureobasidium pullulans, Thekopsora areolata, Phaeococcomyces nigricans, Macrophoma sp., and several Malassezia species. Seasonal differences were observed for community composition, with ascomycetous molds and basidiomycetous yeasts predominating in the winter and spring and Agaricomycetidae basidiomycetes predominating in the fall. The comparison of methods suggested that the cloning, cultivation, and quantitative PCR methods complemented each other, generating a more comprehensive picture of fungal flora than any of the methods would give alone. The current restrictions of the methods are discussed.

Moulds, bacteria and cancer among Finns: an occupational cohort study.

BACKGROUND: Some environmental moulds and bacteria produce carcinogenic toxins. AIM: To study associations between work-related exposure to moulds and bacteria and cancers in Finland. METHODS: A cohort of all economically active Finns in the population census in 1970 were followed-up for 30 million person-years. Subsequent cancer cases were identified through record linkage with the Finnish Cancer Registry. Observed and expected numbers of cancer cases were calculated by occupation, sex, birth cohort and period of observation. Exposures to moulds of agricultural and industrial origin and to bacteria of non-human origin were estimated with the Finnish Job-Exposure Matrix. RESULTS: Men with the highest mould and bacterial exposure had a reduced relative risk for lung cancer (RR 0.7, 95% CI 0.6 to 0.9 for moulds and RR 0.9, 95% CI 0.8 to 1.0 for bacteria). Women in the highest mould and bacterial exposure category had RRs of 3.1 (95% CI 1.0 to 9.2) and 2.6 (95% CI 1.5 to 4.7) for cervical cancer, respectively. The respective RRs for lip cancer were 2.4 (95% CI 1.2 to 5.1) and 1.6 (95% CI 1.2 to 2.2). CONCLUSIONS: Exposures at the investigated concentrations to either moulds or bacteria are unlikely to be major risk factors of cancer, although suggestions of risk increases were observed for some cancer types. It has been suggested previously that the decreased risk for lung cancer is due to the protective effect of endotoxins.

Patients hospitalized abroad as importers of multiresistant bacteria-a cross-sectional study.

OBJECTIVES: The pandemic spread of multidrug-resistant (MDR) bacteria poses a threat to healthcare worldwide, with highest prevalence in indigent regions of the (sub)tropics. As hospitalization constitutes a major risk factor for colonization, infection control management in low-prevalence countries urgently needs background data on patients hospitalized abroad. METHODS: We collected data on 1122 patients who, after hospitalization abroad, were treated at the Helsinki University Hospital between 2010 and 2013. They were screened for methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-PE), vancomycin-resistant enterococci, carbapenemase-producing Enterobacteriaceae (CPE), multiresistant Pseudomonas aeruginosa and multiresistant Acinetobacter baumannii. Risk factors for colonization were explored by multivariate analysis. RESULTS: MDR colonization rates were higher for those hospitalized in the (sub)tropics (55%; 208/377) compared with temperate zones (17%; 125/745). For ESBL-PE the percentages were 50% (190/377) versus 12% (92/745), CPE 3.2% (12/377) versus 0.4% (3/745) and MRSA 6.6% (25/377) versus 2.4% (18/745). Colonization rates proved highest in those returning from South Asia (77.6%; 38/49), followed by those having visited Latin America (60%; 9/16), Africa (60%; 15/25) and East and Southeast Asia (52.5%; 94/179). Destination, interhospital transfer, short time interval to hospitalization, young age, surgical intervention, residence abroad, visiting friends and relatives, and antimicrobial use proved independent risk factors for colonization. CONCLUSIONS: Post-hospitalization colonization rates proved higher in the (sub)tropics than elsewhere; 11% (38/333) of carriers developed an MDR infection. We identified several independent risk factors for contracting MDR bacteria. The data provide a basis for infection control guidelines in low-prevalence countries.

An approach to management of critical indoor air problems in school buildings.

This study was conducted in a school center that had been the focus of intense public concern over 2 years because of suspected mold and health problems. Because several attempts to find solutions to the problem within the community were not satisfactory, outside specialists were needed for support in solving the problem. The study group consisted of experts in civil engineering, indoor mycology, and epidemiology. The studies were conducted in close cooperation with the city administration. Structures at risk were opened, moisture and temperature were measured, and the causes of damage were analyzed. Microbial samples were taken from the air, surfaces, and materials. Health questionnaires were sent to the schoolchildren and personnel. Information on the measurements and their results was released regularly to school employees, students and their parents, and to the media. Repairs were designed on the basis of this information. Moisture damage was caused mainly by difficult moisture conditions at the building site, poor ventilation, and water leaks. Fungal genera (concentrations <200 colony-forming units (cfu)/m(3), <3000 cfu/cm(2)) typical to buildings with mold problems (e.g., Aspergillus versicolor, Eurotium) were collected from the indoor air and surfaces of the school buildings. Where moisture-prone structures were identified and visible signs of damage or elevated moisture content were recorded, the numbers of microbes also were high; thus microbial results from material samples supported the conclusions made in the structural studies. Several irritative and recurrent symptoms were common among the upper secondary and high school students. The prevalence of asthma was high (13%) among the upper secondary school students. During the last 4 years, the incidence of asthma was 3-fold that of the previous 4-year period.

Effect of viability of actinomycete spores on their ability to stimulate production of nitric oxide and reactive oxygen species in RAW264.7 macrophages.

Spores of actinomycetes, mesophilic gram-positive bacteria, isolated from moldy houses, induced the expression of inducible NO-synthase (iNOS) with a subsequent NO-production in RAW264.7 macrophages. No differences were detected between production of nitric oxide (NO) by alive or irradiated spores of different strains of Actinomycetes sp. or Streptomyces sp. Moreover, a significant production of reactive oxygen species (ROS) occurred in the macrophages after their stimulation both by alive and irradiation killed spores of actinomycetes. However, ROS-responses in macrophage induced by dead spores were significantly lower compared to those induced by alive spores. The cytotoxicity of the spores of different actinomycetes differed widely. The production of NO and ROS did not depend directly on the viability of the spores, suggesting an important role for cell wall components in the activation of the cells.

Risk of homicidal behaviour among persons convicted of homicide.

We studied the risk of homicidal behaviour among persons convicted of homicide using an epidemiological method. The results showed that male homicide offenders are at least 10 times more likely to commit a homicide when compared with general male population. This odds ratio was about 150-fold among offenders who had committed at least four previous aggravated violent crimes as well as homicide.

Ammonia, hydrogen sulphide and methyl mercaptides in Finnish municipal sewage plants and pumping stations.

Ammonia, methane, hydrogen sulphide and methyl mercaptides were analyzed in the atmospheres of 16 Finnish municipal waste water treatment plants and in 18 pumping stations. The sulphides were analyzed on site as it was found that, with the standard 0.5-l laminated plastic bags, significant amounts of the vapours were lost from the gas phase. The relative humidity of the sampled air was the most likely cause. Under normal operating conditions, sulphides varied from less than 0.07 to 53 micrograms l-1, with highest concentrations found at the sludge presses. Pumping stations had sulphide vapours from 0.07 to 0.5 microgram l-1. The presence of ammonia (0.007-3.5 micrograms l-1) and methane (0.7-18 micrograms l-1) confirmed the hypothesis of the anaerobic origin of the offensive gases. Although more effective ventilation without scrubbing the emission gases would reduce the health hazard in the plants it would burden the environment.

Exposure to airborne microorganisms and volatile organic compounds in different types of waste handling.

Occupational exposure of workers to airborne microorganisms and volatile organic compounds (VOC) in different types of waste treatment situations was examined during summer time. Microorganisms were collected as stationary samples using a six-stage Andersen impactor, while for VOCs both personal and stationary sampling was conducted. The exposure at the waste handling facility was considerably greater than at landfill sites or in waste collection. The concentrations of viable fungi were maximally 10(5) cfu/m3, and the concentrations of both total culturable bacteria and Gram-negative bacteria exceeded the proposed occupational exposure limit values (OELV), being 10(4) and 10(3) cfu/m3, respectively. Exposure to VOCs in the waste handling facility was three times higher than at the landfill sites, being at highest 3000 microg/m3, considered to be the limit for discomfort. The use of personal protective equipment at work, thorough hand washing and changing clothes after the work shift are strongly recommended in the waste handling facility and the landfill sites.

Analysis of moisture findings in the interior spaces of Finnish housing stock.

A grading system was developed to rate the moisture damage profile of dwellings and to study the relationship between moisture-induced indoor air problems and occupant health. A total of 630 randomly selected houses and apartments, built between 1950 and 1989, were visually inspected. Moisture observations were standardized into three damage levels. Thus, a system to classify the homes into three grades was devised. The two grades of homes associated with the highest levels of damage were graded as index homes. Overall, 51% of the sample had some kind of moisture fault in them and one in every three homes (33%) was classified as an index home. The mean number of damage incidents in the index dwellings varied from 1.4 to 2.6. The mean number of damage incidents in the reference homes was 0.28. Prevalence of index dwellings was significantly higher (p < 0.01) in houses (38%) than in apartments (26%). There was no major difference in the prevalence of index buildings in houses built in any particular decade (30-35%). Moisture was observed in 28% of bathrooms, in 10% of kitchens, and in 17% of other spaces. Indoor relative humidity (RH) levels were low in most homes.

Cytotoxicity, production of reactive oxygen species and cytokines induced by different strains of Stachybotrys sp. from moldy buildings in RAW264.7 macrophages.

The ability of different strains of the fungus Stachybotrys, isolated from mold problem buildings, to induce cytotoxicity and production of important inflammatory mediators, i.e. nitric oxide (NO), reactive oxygen species (ROS), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in RAW264.7 macrophages were studied. Several strains of Stachybotrys sp. stimulated immediate increase in the ROS production and in 24-h exposure caused TNF-α and IL-6 release from these cells. However, none of the strains of Stachybotrys sp. was able to induce the expression of inducible nitric oxide synthase (iNOS) and subsequent production of NO in RAW264.7 cells. Moreover, there were significant differences in their ability to induce cytotoxicity in the macrophages. These results suggest that, in addition to direct cytotoxic effects of most Stachybotrys sp., some strains of Stachybotrys sp. stimulate production of inflammatory mediators, TNF-α and IL-6 which were associated with low cytotoxicity in RAW264.7 macrophages.

Streptomyces spores from mouldy houses induce nitric oxide, TNFα and IL-6 secretion from RAW264.7 macrophage cell line without causing subsequent cell death.

The current view is that only bacterial lipopolysaccharide (LPS) and gamma interferon (IFNγ) are able to alone activate macrophages to secrete nitric oxide (NO), probably a causative agent of cell death. Moreover, some cytokines and gram positive pathogens together with IFNγ induce NO-production. Surprisingly, spores of Streptomyces sp., which are mesophilic gram-positive bacteria found in mouldy houses, stimulated RAW264.7 macrophages to produce pro-inflammatory cytokines, tumor necrosis factor alpha (TNFα) and interleukin-6 (IL-6), and induced the expression of inducible NO-synthase (iNOS) with a subsequent NO-production. However, the Streptomyces spores did not kill NO-producing macrophages, as did both LPS and gram negative bacteria Pseudomonas fluorescens, strong inducers of cytokine- and NO-production. These results imply that Streptomyces sp., induced cytokine and NO-secretion, may play a role in the responses evoked by exposure to these microbes. Moreover, factors other than, or in addition to NO, are necessary for cytotoxicity in murine macrophages.