Fungal contamination in swine: a potential occupational health threat

Poor air quality in a pig-confinement building may potentially place farmers at higher health risk than other workers for exposure to airborne pollutants that may reach infectious levels. The aim of this study was to assess worker exposure to fungi in indoor environments in Portuguese swine buildings. Air samples from 7 swine farms were collected at a flow rate of 140 L/min, at 1 m height, onto malt extract agar supplemented with chloramphenicol (MEA). Surfaces samples of the same indoor sites were obtained by swabbing the surfaces. Samples from the floor covering were also collected from four of seven swine farms. All collected samples were incubated at 27°C for 5-7 days. After lab processing and incubation of obtained samples, quantitative colony-forming units (CFU)/m(3), CFU/cm(2), and CFU/g and qualitative results were determined with identification of isolated fungal species. Aspergillus versicolor was the most frequent species found in air (21%), followed by Scopulariopsis brevicaulis (17%) and Penicillium sp. (14%). Aspergillus versicolor was also the most frequent species noted on surfaces (26.6%), followed by Cladosporium sp. (22.4%) and Scopulariopsis brevicaulis (17.5%). Chrysosporium was the most frequently found genera in the new floor covering (38.5%), while Mucor was the most prevalent genera (25.1%) in used floor covering. Our findings corroborate a potential occupational health threat due to fungi exposure and suggest the need for a preventive strategy.

Occupational exposure to Aspergillus by swine and poultry farm workers in Portugal

Aspergillus is among a growing list of allergens that aggravate asthmatic responses. Significant pulmonary pathology is associated with Aspergillus-induced allergic and asthmatic lung disease. Environments with high levels of exposure to fungi are found in animal production facilities such as for swine and poultry, and farmers working with these are at increased risk for occupational respiratory diseases. Seven Portuguese poultry and seven swine farms were analyzed in order to estimate the prevalence, amount, and distribution of Aspergillus species, as well as to determine the presence of clinical symptoms associated with asthma and other allergy diseases in these highly contaminated settings. From the collected fungal isolates (699), an average incidence of 22% Aspergillus was detected in poultry farms, while the prevalence at swine farms was 14%. The most frequently isolated Aspergillus species were A. versicolor, A. flavus, and A. fumigatus. In poultry farms, A. flavus presented the highest level of airborne spores (>2000 CFU/m3), whereas in swine farms the highest was A. versicolor, with an incidence fourfold greater higher than the other mentioned species. Eighty workers in these settings were analyzed, ranging in age from 17 to 93 yr. The potentially hazardous exposure of poultry workers to mold allergens using sensitization markers was evaluated. Although no significant positive association was found between fungal contamination and sensitization to fungal antigens, a high incidence of respiratory symptoms in professionals without asthma was observed, namely, wheezing associated with dyspnea (23.8%) and dyspnea after strenuous activities (12.3%), suggesting underdiagnosed respiratory disturbances. Further, 32.5% of all exposed workers noted an improvement of respiratory ability during resting and holidays. From all the analyzed workers, seven were previously diagnosed with asthma and four reported the first attack after the age of 40 yr, which may be associated with their occupational exposure. Some of the fungi, namely, the Aspergillus species detected in this study, are known to induce hypersensitivity reactions in humans. This study confirmed the presence and distribution of Aspergillus in Portuguese poultry and swine farms, suggesting a possible occupational health problem and raising the need for preventive and protective measures to apply to avoid exposure in both occupational settings.

Effects of fungal contamination on respiratory symptoms of poultry workers

Exposure to certain fungi (molds) can cause human illness by 3 specific mechanisms: generation of a harmful immune response, direct infection by the organism or/and toxic-irritant effects from mold byproducts. Moulds are considered central elements in daily exposure of poultry workers and can be the cause of an increased risk of occupational respiratory diseases, like allergic and non-allergic rhinitis and asthma.

Exposure to particles in Portuguese swine production

Several studies have shown that human exposures to airbome dust and microorganisms, such as bacteria and fungi, can cause respiratory diseases. Agricultural workers have been found to be at high risk of exposures to airborne particles. From a human health perspective dust exposure in pig farming is the most important risk because of the large number of workers needed in pig production and the increasing number of working hours inside enclosed buildings. In the pig buildings, particulate matters like dust play a role in not only deteriorating indoor air quality but also can cause an adverse health effect on workers. Generally, dust is recognized to adsorb and transport odorous compounds and biological agents. The aim of this study was to determine particles contamination in 7 swine farms located in Lisbon district, Portugal.

Occupational exposure to toxigenic Aspergillus versicolor in Portuguese swines

Biological factors associated with airbome dust are the most important hazards in pig buildings and include allergenic and/or toxic compounds, as well as infectious agents such as fungi and their metabolites, like mycotoxins. Inhalation of such agents can be a potential occupationai treat. Exposure of workers from swine confinement buildings to respiratory hazards has been reported elsewhere in Europe, Asia and America. Analogous data has not been reported for Portugal and this omission has hindered the development of policies in the area of occupational health and farm safety. Aspergilius versicolor is known as being the major producer of the hepatotoxic and carcinogenic mycotoxin sterigmatocystin. The toxicity of this mycotoxin is manifested primarily in liver and kidney. This study aimed to determine occupational exposure treat due to fungal contamination caused by A. versicolor in seven Portuguese swine.

Fungal contamination: comparation between Portuguese poultries and swines

Animal confinement tends to increase the overall microbial load in the production environment caused by high amounts of feed and organic residuals (manure and wastewater) present in those environments. The number of animais and the handling and management required to work in these settings also contribute to enhance that microbial ioad. Animal housing typically exposes workers to substantial concentrations of bioaerosols, such as fungi and their metabolites. Therefore, agricultural workers, and especially pig and poultry farmers, are at increased risk of occupational respiratory diseases. Exposure to bioaerosols in poultries and swines may vary depending upon the stage of the animals' growth, density, manure management procedures, litter type and used floor coverage, among others. Gathering temporal information about the quantity and the composition of fungal load is necessary to better understand the relationship between these factors and adverse health symptoms of workers. This study aimed to characterize and compare fungal contamination between these two different settings.

Fungi, MVOCs and dust exposure assessment in poultry production

Agricultural workers especially poultry farmers, are at increased risk of occupational respiratory diseases. In poultry production besides fungi microbial volatile organic compounds (MVOCs) are also present due to compounds released during fungal metabolism. Dust is also one of the risk factors present in animal housing and is comprised by poultry residues, fungi and feathers. A study was developed aiming to assess occupational exposure to fungi, MVOCs and dust in seven poultry units located in Portugal.

Effects of fungal contamination on respiratory symptoms of poultry workers

Moulds are considered central elements in daily exposure of poultry workers and can be the cause of an increased risk of occupational respiratory diseases, like allergic and non-allergic rhinitis and asthma. The objective is to evaluate the exposure to different species of moulds in poultries and relate them with respiratory symptoms in poultry workers. Seven Portuguese poultries were analyzed in order to assess air fungal contamination, as well as to evaluate the existence of clinical symptoms associated with asthma and other allergy diseases by European Community Respiratory Health Survey questionnaire.

Occupational exposure to Aflatoxin B1 in swine production and possible contamination sources

Although the adverse health consequences of ingestion of food contaminated with aflatoxin B1 (AFB1) are known, relatively few studies are available on the adverse effects of exposure in occupational settings. Taking this into consideration, our study was developed aiming to elucidate the possible effects of occupational exposure to AFB1 in Portuguese swine production facilities using a specific biomarker to assess exposure to AFB1. In total, 28 workers participated in this study, providing blood samples, and a control group (n  = 30) was composed of subjects without any type of agricultural activity. Fungal contamination was also studied by conventional methods through air, surfaces, and new and used floor coverage. Twenty-one workers (75%) showed detectable levels of AFB1 with values ranging from <1 ng/ml to 8.94 ng/ml and with a mean value of 1.91 ± 1.68 ng/ml. In the control group, the AFB1 values were all below 1 ng/ml. Twelve different Aspergillus species were identified. Aspergillus versicolor presented the highest airborne spore counts (3210 CFU/m3) and was also detected in higher values in surfaces (>300 CFU/cm2). Data indicate that exposure to AFB1 occurs in swine barns, and this site serves as a contamination source in an occupational setting.

Occupational exposure to aflatoxin B1: the case of poultry and swine production

Although there is an abundance of literature concerning the ingestion of food contaminated with aflatoxin B1 (AFB1), only a small number of studies explore mycotoxin exposure in occupational settings. Taking this into consideration, our study was developed with the intention of elucidating whether there is occupational exposure to AFB1 in Portuguese poultry and swine production facilities. A specific biomarker was used to assess exposure to AFB1. A total of 45 workers (34 from poultry farms; 11 from swine production facilities) participated in this study, providing blood samples. Additionally, a control group (n=30) composed of subjects without any type of contact with agricultural activity was considered. All participants signed a consent form and were provided with the study protocol. Eighteen poultry workers (58.6%) and six workers from the swine production facilities (54.5%) showed detectable levels of AFB1. In the control group, the AFB1 values were all below 1 ng/ml. No significant differences in AFB1 levels in serum between workers from poultry and swine farms were found. Poultry workers, however, showed the highest serum levels and a significant statistical difference between this group and the control group was found. Results suggest that exposure to AFB1 by inhalation occurs in both occupational settings representing an additional risk that needs to be recognised, assessed and prevented.

Exposure to particles and fungi in Portuguese swine production

Versão preprint.

Accessing indoor fungal contamination using conventional and molecular methods in Portuguese poultries

Epidemiological studies showed increased prevalence of respiratory symptoms and adverse changes in pulmonary function parameters in poultry workers, corroborating the increased exposure to risk factors, such as fungal load and their metabolites. This study aimed to determine the occupational exposure threat due to fungal contamination caused by the toxigenic isolates belonging to the complex of the species of Aspergillus flavus and also isolates fromAspergillus fumigatus species complex. The study was carried out in seven Portuguese poultries, using cultural and molecularmethodologies. For conventional/cultural methods, air, surfaces, and litter samples were collected by impaction method using the Millipore Air Sampler. For the molecular analysis, air samples were collected by impinger method using the Coriolis μ air sampler. After DNA extraction, samples were analyzed by real-time PCR using specific primers and probes for toxigenic strains of the Aspergillus flavus complex and for detection of isolates from Aspergillus fumigatus complex. Through conventional methods, and among the Aspergillus genus, different prevalences were detected regarding the presence of Aspergillus flavus and Aspergillus fumigatus species complexes, namely: 74.5 versus 1.0% in the air samples, 24.0 versus 16.0% in the surfaces, 0 versus 32.6% in new litter, and 9.9 versus 15.9%in used litter. Through molecular biology, we were able to detect the presence of aflatoxigenic strains in pavilions in which Aspergillus flavus did not grow in culture. Aspergillus fumigatus was only found in one indoor air sample by conventional methods. Using molecular methodologies, however, Aspergillus fumigatus complex was detected in seven indoor samples from three different poultry units. The characterization of fungal contamination caused by Aspergillus flavus and Aspergillus fumigatus raises the concern of occupational threat not only due to the detected fungal load but also because of the toxigenic potential of these species.

Occupational exposure to particulate matter and respiratory symptoms in Portuguese swine barn workers

Certain environmental conditions in animal and plant production have been associated with increased frequency in respiratory illnesses, including asthma, chronic bronchitis, and hypersensitivity pneumonitis, in farmers occupationally exposed in swine production. The aim of this study was to characterize particulate matter (PM) contamination in seven Portuguese swine farms and determine the existence of clinical symptoms associated with asthma and other allergy diseases, utilizing the European Community Respiratory Health Survey questionnaire. Environmental assessments were performed with portable direct-reading equipment, and PM contamination including five different sizes (PM0.5, PM1.0, PM2.5, PM5.0, PM10) was determined. The distribution of particle size showed the same trend in all swine farms, with high concentrations of particles with PM5 and PM10. Results from the questionnaire indicated a trend such that subjects with diagnosis of asthma were exposed to higher concentrations of PM with larger size (PM2.5, PM5, and PM10) while subjects with sneezing, runny nose, or stuffy nose without a cold or flu were exposed to higher concentrations of PM with smaller size (PM0.5 and PM1). Data indicate that inhalation of PM in swine farm workers is associated with increased frequency of respiratory illnesses.

Occupational exposure to toxigenic fungi from Aspergillus flavus complex

Bioaerosols are mainly composed of fungal particles, bacteria and plant spores, being fungi responsible for the release of VOCs and micotoxins into indoor environments. Aspergillus flavus is a common opportunistic pathogen causing human infections and is involved in the production of aflatoxin and other secondary metabolites associated with toxic and allergic reactions. Poultry workers are exposed to high concentrations of fungi and are therefore more prone to develop associated pathologies. To evaluate occupational exposure of the workers to Aspergillus flavus and aflatoxins, six animal production facilities were selected, including 10 buildings, from which indoor air samples and outdoor reference samples were obtained. Twenty-five duplicate samples were collected by two methodologies: impactation onto malt extract agar of 25L air samples using a Millipore Air Tester were used to evaluate quantitative (CFU/m3) and qualitative (species identification, whenever possible) sample composition; 300 L air samples collected with the Coriolis Air Sampler into phosphate–saline buffer were used to isolate DNA, following molecular identification of Aspergillus section flavi using nor-1 specific primers by real-time PCR.

Exposure to fungi in dockers

Several activities are ensured by dockers increase occupational exposure to several risk factors. being one of them the fungal burden from the load. In this study we aim at characterizing fungal contamination in one warehouse that storage sugar cane from a ship, and also in one crane cabinet that unload the same sugar cane from the ship. Air samples were collected from the warehouse and from inside the crane cabinet. An outdoor sample was also collected, from each sampling site, and regarding as reference. Sampling volume was selected depending in the contamination expected and the air samples were collect through an impaction method in a flow rate of 140 L/min onto malt extract agar (MEA) supplemented with chloramphenicol (0.05%), using the Millipore air Tester (Millipore). Surfaces samples from the warehouse were collected by swabbing the surfaces of the same indoor sites, using a 10 by 10cm square stencil according to the International Standard ISO 18593 (2004). The obtained swabs were then plated onto MEA. All the collected samples were incubated at 27ºC for 5 to 7 days. After laboratory processing and incubation of the collected samples, quantitative (colony-forming units - CFU/m3 and CFU/m2) and qualitative results were obtained with identification of the isolated fungal species. Aspergillus fumigatus present the highest fungal load and WHO guideline was overcome in both indoor sampling sites. The results obtained in this study highlight the need to know better the exposure burden from dockers, and specifically to fungi contamination.