Potential poultry and meat products contamination by aflatoxin B1 due to fungal presence in Portuguese poultry units

The impact of mycotoxins on human and animal health is well recognized. Aflatoxin B1 (AFB1) is by far the most prevalent and the most potent natural carcinogen and is usually the major aflatoxin produced by toxigenic fungal strains. Data available, points to an increasing frequency of poultry feed contamination by aflatoxins. Since aflatoxin residues may accumulate in body tissues, this represents a high risk to human health. Samples from commercial poultry birds have already presented detectable levels of aflatoxin in liver. A descriptive study was developed in order to assess fungal contamination by species from Aspergillus flavus complex in seven Portuguese poultry units. Air fungal contamination was studied by conventional and molecular methods. Air, litter and surfaces samples were collected. To apply molecular methods, air samples of 300L were collected using the Coriolis μ air sampler (Bertin Technologies), at 300 L/min airflow rate. For conventional methodologies, all the collected samples were incubated at 27ºC for five to seven days. Through conventional methods, Aspergillus flavus was the third fungal species (7%) most frequently found in 27 indoor air samples analysed and the most commonly isolated species (75%) in air samples containing only the Aspergillus genus...

Ventilation influence in occupational exposure to fungi and volatile organic compounds: poultry case

Introduction - In poultry houses, large-scale production has led to increased bird densities within buildings. Such high densities of animals kept within confined spaces are a source of human health problems related to occupational organic dust exposure. This organic dust is composed of both non-viable particles and viable particulate matter (also called bioaerosols). Bioaerosols are comprised by airborne bacteria, fungi, viruses and their by-products, endotoxins and mycotoxins. Exposure to fungi in broiler houses may vary depending upon the applied ventilation system. Ventilation can be an important resource in order to reduce air contamination in these type of settings. Nevertheless, some concerns regarding costs, sensitivity of the animal species to temperature differences, and also the type of building used define which type of ventilation is used. Aim of the study - A descriptive study was developed in one poultry unit aiming to assess occupational fungal and volatile organic compounds (VOCs) exposure.

Fungal contamination of poultry litter: a public health problem

Although numerous studies have been conducted on microbial contaminants associated with various stages related to poultry and meat products processing, only a few reported on fungal contamination of poultry litter. The goals of this study were to (1) characterize litter fungal contamination and (2) report the incidence of keratinophilic and toxigenic fungi presence. Seven fresh and 14 aged litter samples were collected from 7 poultry farms. In addition, 27 air samples of 25 litters were also collected through impaction method, and after laboratory processing and incubation of collected samples, quantitative colony-forming units (CFU/m3) and qualitative results were obtained. Twelve different fungal species were detected in fresh litter and Penicillium was the most frequent genus found (59.9%), followed by Alternaria (17.8%), Cladosporium (7.1%), and Aspergillus (5.7%). With respect to aged litter, 19 different fungal species were detected, with Penicillium sp. the most frequently isolated (42.3%), followed by Scopulariopsis sp. (38.3%), Trichosporon sp. (8.8%), and Aspergillus sp. (5.5%). A significant positive correlation was found between litter fungal contamination (CFU/g) and air fungal contamination (CFU/m3). Litter fungal quantification and species identification have important implications in the evaluation of potential adverse health risks to exposed workers and animals. Spreading of poultry litter in agricultural fields is a potential public health concern, since keratinophilic (Scopulariopsis and Fusarium genus) as well as toxigenic fungi (Aspergillus, Fusarium, and Penicillium genus) were isolated.

Influence of ventilation type in volatile organic compounds exposure: poultry case

Agricultural workers especially poultry farmers are at increased risk of occupational respiratory diseases. Epidemiological studies showed increased prevalence of respiratory symptoms and adverse changes in pulmonary function parameters in poultry workers. In poultry production volatile organic compounds (VOCs) presence can be due to some compounds produced by molds that are volatile and are released directly into the air. These are known as microbial volatile organic compounds (MVOCs). Because these compounds often have strong and/or unpleasant odors, they can be the source of odors associated with molds. MVOC's are products of the microorganisms primary and secondary metabolism and are composed of low molecular weight alcohols, aldehydes, amines, ketones, terpenes, aromatic and chlorinated hydrocarbons, and sulfur-based compounds, all of which are variations of carbon-based molecules.

Assessment of toxigenic fungi in poultry feed

Introduction - Feed supplies the necessary nutrients for the growth of healthy animals, which are a part of the human diet. The presence of toxigenic fungi in animal feed such as Aspergillus spp. may contribute to 1) the loss of nutritional value of feedstuff, since fungi will assimilate the most readily available nutrients present in the feed, and 2) the development of mycotoxicoses and chronic conditions, which can raise economic issues due to animal disease and contamination of animal derived products. Aim of the study - The goal of this work was to evaluate the incidence of Aspergilli, particularly from the Circumdati, Flavi and Fumigati sections, through real-time quantitative PCR (qPCR) in 11 feed samples.

Development of a penetration test for timber impregnation products for use in old buildings

This paper studies the application of commercial biocides to old maritime pine timber structures (Pinus pinaster Ait.) that have previously been impregnated with other products. A method was developed in the laboratory to be used in situ to determine the impregnation depth achieved by a new generation biocide product applied to timber from an old building. This timber had once been treated with an unknown product difficult to characterize without extensive analysis. The test was initially developed in laboratory conditions and later tested on elements of the roof structure of an 18th century building. In both cases the results were promising and mutually consistent with penetration depths for some treatments reaching 2.0 cm. The application in situ proved the tests viability and simplicity of execution giving a clear indication on the feasibility of possible re-treatments.

Possíveis implicações da contaminação fúngica num aviário

Introdução – Apesar de em Portugal se verificar o aumento da indústria da produção de aves para consumo humano, apenas alguns estudos incidem sobre a qualidade do ar interior e as implicações da sua degradação. Objectivos – Descrever a contaminação fúngica num aviário, analisar possíveis associações com a temperatura ambiente e a humidade relativa e o possível impacto na saúde dos consumidores e trabalhadores desta unidade. Métodos – Foi desenvolvido um estudo descritivo para avaliar a contaminação fúngica num aviário. Colheram‑se 5 amostras de ar de 100 litros através do método de compactação e 4 amostras de superfícies, utilizando a técnica da zaragatoa e um quadrado de 10 cm de lado de metal. Simultaneamente, os parâmetros ambientais – temperatura ambiente e humidade relativa – também foram medidos. Resultados – Foram identificadas vinte espécies de fungos no ar, sendo os seguintes os quatro géneros mais comummente isolados: Cladosporium (40,5%), Alternaria (10,8%), Chrysosporium e Aspergillus (6,8%). Nas superfícies, 21 espécies de fungos foram identificadas, sendo os 4 géneros mais identificados Penicillium (51,8%), Cladosporium (25,4%), Alternaria (6,1%) e Aspergillus (4,2%). Importa referir o facto de Aspergillus flavus, também isolado no ar, ser reconhecido como produtor de micotoxinas (aflatoxina) e Aspergillus fumigatus, uma das espécies isoladas no ar e superfícies, ser capaz de causar aspergilose grave ou fatal. Não se verificou relação significativa (p> 0,05) entre a contaminação fúngica e as variáveis ambientais. Conclusão – Caracterizou‑se a distribuição fúngica no ar e superfícies do aviário e analisou‑se a possível influência das variáveis ambientais. Foi reconhecido um potencial problema de Saúde Pública devido à contaminação fúngica e à possível produção de micotoxinas com a eventual contaminação dos produtos alimentares. A contaminação fúngica, particularmente causada pelo Aspergillus fumigatus, e a possível presença de micotoxinas no ar, devem ser encaradas também como fatores de risco neste contexto ocupacional. ABSTRACT - Background – Although there is an increasingly industry that produce whole chickens for domestic consumption in Portugal, only few investigations have reported on the indoor air of these plants and the consequences of their degradation. Objectives – Describe one poultry environmental fungal contamination analyse possible associations between temperature and relative humidity and its possible impact on the health of consumers and of the poultry workers. Methods – A descriptive study was developed to monitor one poultry fungal contamination. Five air samples of 100 litres through impaction method were collected and 4 swab samples from surfaces were also collected using a 10 cm square of metal. Simultaneously, environmental parameters – temperature and relative humidity – were also measured. Results – Twenty species of fungi in air were identified, being the 4 most commonly isolated the following genera: Cladosporium (40.5%), Alternaria (10.8%), Chrysosporium and Aspergillus (6.8%). In surfaces, 21 species of fungi were identified, being the 4 genera more identified Penicillium (51.8%), Cladosporium (25.4%), Alternaria (6.1%) and Aspergillus (4.2%). In addition, Aspergillus flavus also isolated in the poultry air is a well‑known producer of potent mycotoxins (aflatoxin), and Aspergillus fumigatus, one of the species isolated in air and surfaces, is capable of causing severe or fatal aspergillosis. There was no significant relationship (p>0,05) between fungal contamination and environmental variables. Conclusions – Was characterized fungal distribution in poultry air and surfaces and analyzed the association of environmental variables. It was recognized the Public Health problem because of fungal contamination and also due to probable mycotoxins production with the possible contamination of food products. Fungal contamination, particularly due to the presence of Aspergillus fumigatus and also the possible presence of mycotoxins in the air, should be seen as risk factor in this occupational setting.

Occupational exposure to aflatoxin (AFB1) in poultry production

Aflatoxin B1 (AFB1) has been recognized to cause cancer in the humans liver. Epidemiological and laboratory evidence also point towards the respiratory system as target for carcinogenesis. Exposure occurs mainly through food chain but inhalation represent an additional route of exposure. Agricultural workers have the greatest risk of occupational exposure due to their exposition to airborne aflatoxin through inhalation of dust.

Air contaminants in animal production: the poultry case

A descriptive study was developed in order to assess air contamination caused by fungi and particles in seven poultry units. Twenty seven air samples of 25 litters were collected through impaction method. Air sampling and particle concentration measurement were performed in the pavilions’ interior and also outside premises, since this was the place regarded as reference. Simultaneously, temperature and relative humidity were also registered. Regarding fungal load in the air from the seven poultry farms, the highest value obtained was 24040 CFU/m3 and the lowest was 320 CFU/m3. Twenty eight species/genera of fungi were identified, being Scopulariopsis brevicaulis (39.0%) the most commonly isolated species and Rhizopus sp. (30.0%) the most commonly isolated genus. From the Aspergillus genus, Aspergillus flavus (74.5%) was the most frequently detected species. There was a significant correlation (r=0.487; p=0.014) between temperature and the level of fungal contamination (CFU/m3). Considering contamination caused by particles, in this study, particles with larger dimensions (PM5.0 and PM10) have higher concentrations. There was also a significant correlation between relative humidity and concentration of smaller particles namely, PM0.5 (r=0.438; p=0.025) and PM1.0 (r=0.537; p=0.005). Characterizing typical exposure levels to these contaminants in this specific occupational setting is required to allow a more detailed risk assessment analysis and to set exposure limits to protect workers’ health.

Comparison of indoor and outdoor fungi and particles in poultry units

A descriptive study was developed in order to compare indoor and outdoor air contamination caused by fungi and particles in seven poultry units. Twenty eight air samples of 25 litters were collected through the impaction method on malt extract agar. Air sampling and particles concentration measurement were done in the interior and also outside premises of the poultries’ pavilions. Regarding the fungal load in the air, indoor concentration of mold was higher than outside air in six poultry units. Twenty eight species / genera of fungi were identified indoor, being Scopulariopsis brevicaulis (40.5%) the most commonly isolated species and Rhizopus sp. (30.0%) the most commonly isolated genus. Concerning outdoor, eighteen species/genera of fungi were isolated, being Scopulariopsis brevicaulis (62.6%) also the most isolated. All the poultry farms analyzed presented indoor fungi different from the ones identified outdoors. Regarding particles’ contamination, PM2.5, PM5.0 and PM10 had a statistically significant difference (Mann-Whitney U test) between the inside and outside of the pavilions, with the inside more contaminated (p=.006; p=.005; p=.005, respectively). The analyzed poultry units are potential reservoirs of substantial amounts of fungi and particles and could therefore free them in the atmospheric air. The developed study showed that indoor air was more contaminated than outdoors, and this can result in emission of potentially pathogenic fungi and particles via aerosols from poultry units to the environment, which may post a considerable risk to public health and contribute to environmental pollution.

Occupational exposure to aflatoxin (AFB1) in poultry production

Aflatoxin B1 (AFB1) has been recognized to produce cancer in human liver. In addition, epidemiological and laboratory studies demonstrated that the respiratory system was a target for AFB1. Exposure occurs predominantly through the food chain, but inhalation represents an additional route of exposure. The present study aimed to examine AFB1 exposure among poultry workers in Portugal. Blood samples were collected from a total of 31 poultry workers from six poultry farms. In addition, a control group (n = 30) was included comprised of workers who undertook administrative tasks. Measurement of AFB1 in serum was performed by enzyme-linked immunosorbent assay (ELISA). For examining fungi contamination, air samples were collected through an impaction method. Air sampling was obtained in pavilion interior and outside the premises, since this was the place regarded as the reference location. Using molecular methods, toxicogenic strains (aflatoxin-producing) were investigated within the group of species belonging to Aspergillus flavus complex. Eighteen poultry workers (59%) had detectable levels of AFB1 with values ranging from <1 ng/ml to4.23 ng/ml and with a mean value of 2 ± 0.98ng/ml. AFB1 was not detected in the serum sampled from any of the controls. Aspergillus flavus was the fungal species third most frequently found in the indoor air samples analyzed (7.2%) and was the most frequently isolated species in air samples containing only Aspergillus genus (74.5%). The presence of aflatoxigenic strains was only confirmed in outdoor air samples from one of the units, indicating the presence of a source inside the building in at least one case. Data indicate that AFB1 inhalation represents an additional risk in this occupational setting that needs to be recognized, assessed, and prevented.

Biological monitoring of aflatoxin (AFB1) in workers of swine and poultry production

Although a great body of literature exists concerning the ingestion of food contaminated with aflatoxin, there are still few studies regarding mycotoxin inhalation in occupational settings. Since mycotoxins are relatively non-volatile, inhalation exposure is cause by inhalation of airborne fungal particulates or fungi-contaminated substrates that contain aflatoxin. We intend to know if there is occupational exposure to aflatoxin in Portuguese poultry and swine production. A total of 19 individuals (11 swine; 8 poultry) agreed and provided blood samples during the course of this investigation. Measurement of AFB1 was performed by ELISA. The samples were treated with pronase (Merck), wash in a Column C18 and purification was made with immunoaffinity columns (R.biopharma), specific for AFB1. It was applied statistical test (Mann-Whitney) to verified statistical difference in AFB1 results between the two settings. Results varied with concentrations from Poultry: poultry workers have higher aflatoxin values, probably because poultry activities are promoter of particles exposure. Only women’s in both settings have results

Exposure to dust in poultry: the importance of task differences for detailed exposure assessment

Dust is a complex mixture of particles of organic and inorganic origin and different gases absorbed in aerosol droplets. In a poultry unit include dried faecal matter and urine, skin flakes, ammonia, carbon dioxide, pollens, feed and litter particles, feathers, grain mites, fungi spores, bacteria, viruses and their constituents. Dust particles vary in size and differentiation between particle size fractions is important in health studies in order to quantify penetration within the respiratory system. A descriptive study was developed in order to assess exposure to particles in a poultry unit during different operations, namely routine examination and floor turn over. Direct-reading equipment was used (Lighthouse, model 3016 IAQ). Particle measurement was performed in 5 different sizes (PM0.5; PM1.0; PM2.5; PM5.0; PM10). The chemical composition of poultry litter was also determined by neutron activation analysis. Normally, the litter of poultry pavilions is turned over weekly and it was during this operation that the higher exposure of particles was observed. In all the tasks considered PM5.0 and PM10.0 were the sizes with higher concentrations values. PM10 is what turns out to have higher values and PM0.5 the lowest values. The chemical element with the highest concentration was Mg (5.7E6 mg.kg-1), followed by K (1.5E4 mg.kg-1), Ca (4.8E3 mg.kg-1), Na (1.7E3 mg.kg-1), Fe (2.1E2 mg.kg-1) and Zn (4.2E1 mg.kg-1). This high presence of particles in the respirable range (<5–7μm) means that poultry dust particles can penetrate into the gas exchange region of the lung. Larger particles (PM10) present a range of concentrations from 5.3E5 and 3.0E6 mg/m3.

Occupational exposure to poultry dust and effects on the respiratory system in workers

Farmers are occupationally exposed to many respiratory hazards at work and display higher rates of asthma and respiratory symptoms than other workers. Dust is one of the components present in poultry production that increases risk of adverse respiratory disease occurrence. Dust originates from poultry residues, molds, and feathers and is biologically active as it contains microorganisms. Exposure to dust is known to produce a variety of clinical responses, including asthma, chronic bronchitis, chronic airways obstructive disease (COPD), allergic alveolitis, and organic dust toxic syndrome (ODTS). A study was developed to determine particle contamination in seven poultry farms and correlate this with prevalence rate of respiratory defects and record by means of a questionnaire the presence of clinical symptoms associated with asthma and other allergy diseases by European Community Respiratory Health Survey. Poultry farm dust contamination was found to contain higher concentrations of particulate matter (PM) PM5 and PM10. Prevalence rate of obstructive pulmonary disorders was higher in individuals with longer exposure regardless of smoking status. In addition, a high prevalence for asthmatic (42.5%) and nasal (51.1%) symptoms was noted in poultry workers. Data thus show that poultry farm workers are more prone to suffer from respiratory ailments and this may be attributed to higher concentrations of PM found in the dust. Intervention programs aimed at reducing exposure to dust will ameliorate occupational working conditions and enhance the health of workers.

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.