Polish Norm PN-89/Z-04111/02 and PN-89/Z-04111/03 (
1. Introduction
Intensive poultry production, implying large densities of animals in small areas, is a significant source of air pollution which may constitute a considerable health hazard to the birds, farmers and those living in the proximity of the farm (Lonc & Plewa, 2009). On the other hand, the spread of bioaerosols on the outside of animal housing may result in local or even more extensive environmental pollution (Bakutis et al., 2004).
Under commercial production the airborne particles will contain a mixture of biological material from a range of sources. The chickens produce large amounts of dust as a result of epithelial desquamation, as well as from feed, manure, faeces and litter (Matković et al., 2009). This dust consists of a variety of airborne particles of biological origin, i.e. bacteria, fungi, endotoxins (lipopolysaccharide, LPS) of Gram-negative bacteria, 1.3-beta-glucan of fungi, fungal spores and mycelium fragments. Hence, a more descriptive term for these airborne particles is bioaerosol in which the microorganisms can occur either as liquid droplets or as dry particles [Dutkiewicz, 1987; Matković et al., 2009; Nevalainen, 2007]. In specific conditions, bioaerosols may show pathogenic, toxic or allergy-causing effects. The particles in a bioaerosol are generally 0.3 to 100 µm in diameter; however, the respirable size fraction of 1 to 10 µm is of primary concern. Bioaerosols, ranging in size from 1.0 to 5.0 µm, generally remain in the air, whereas larger particles are deposited on surfaces (Srikanth et al., 2008).
Bioaerosol may contain representatives of Gram-positive bacteria:
As with bacteria, the fungi present in the poultry dust bioaerosols may be derived from soil, dust feed and litter, but to a lesser extent from the birds themselves. Fungi are ubiquitous in all atmospheres. In general, both outdoor and indoor atmospheres are dominated by representatives of the genera
Literature data usually pertain to the air biopollutant concentration inside the poultry houses. Much less is known about the relationships between the indoor and outdoor biological pollution, as well as about the spreading of indoor bioaerosols in the surroundings of the farms.
The aim of the study was to assess the influence of microbiological air contamination in the intensive poultry breeding, both inside and outside farms. The comparative quantity and quality analysis concerns bacteria as well as fungi isolated from the air samples taken during two seasons.
2. Materials and methods
Seasonal sampling was conducted in the summer of 2009 and spring of 2010 in two (I and II) poultry houses on family farms located near Wrocław in Lower Silesia, Poland (Fig. 1.)
Both farms were accommodated to 18 000 and 23 000 broilers, with the density of 16-17 chicken per square meter. The broilers were kept on the rye straw deep litter in buildings equipped with mechanical ventilation (inlet and outlet ventilators), heating with a central thermogen and artificial lighting with regularly distributed bulbs.
Air samples were taken using a MAS-100 air sampler (Merck KgaA, Darmstadt, Germany) which is representative of the new generation impactor samplers and is frequently used for indoor and outdoor sampling. These instruments are based on the principles described by Andersen and aspirate air through a perforated plate, which results in impaction of particles from an airstream onto the surface of agar medium. The speed of air flow through the sampler was about 11 m/s, air volumes were 5-200 litres (depending on the expected contamination level) and the sampling rate was 100 l/min. Indoor and outdoor samples were collected in the poultry biozone during the fattening period. The biopollutants were determined on the basis of airborne bacteria and fungi. The samples for each group of bacteria and fungi were taken at the central point of poultry houses 1.3 m from the ground level. The emission level outside the farming objects was determined similarly, i.e. 1.3 m with sampling points situated 10 m, 50 m and 100 m away from the farming buildings. At the same time both humidity and temperature were measured with a termohigrometer (Label).
Microbiological studies of the air samples were used to determine the number of mesophilic bacteria,
(BioMérieux) plates were inoculated for culturing and counting
where:
N = 400 (number of holes in perforated lid of the sampler)
r - number of CFU counted on Petri dish
Pr - statistically corrected total count of bacteria/moulds in tested air volume
Bacterial species were identified on the basis of gram staining, microscopic morphology, oxidase and coagulase activity, catalase test results and metabolic properties according to standard procedures described in Bergey’s Manual of Determinative Bacteriology (2001). The following commercial systems were used: API 20E (BioMérieux, France) for enteric gram-negative organisms; API 20 NE (BioMérieux) for fastidious and nonfermenting gram-negative organisms; API Staph (BioMérieux) for gram-positive staphylococci, API 20C AUX (BioMérieux) for identification of yeasts, Slidex - Strepto Kit (BioMérieux) for the identification of Lancefield A,B,C,D,F et G group antigens of streptococci and Slidex Staph Plus (BioMérieux) to detect clumping factor, protein A and group-specific antigen bound to the
3. Results. Quantitative and qualitative relationships between indoor and outdoor microflora examined in summer and spring
The studies were carried out in the summer of 2009 and in the spring of 2010, when the temperature of atmospheric air ranged between 15.20C and + 24.50C; the inside temperature in the poultry houses varied from 220C to 270C. Indoor relative air humidity was about 70-85 %, outdoor ca. 38-82%.
For both poultry houses, the indoor concentration of bacteria and moulds were always higher compared with the outdoor concentration at distance 10 m, 50 m and 100 m from the poultry houses. The number of microorganisms (as CFU/m3) in the atmospheric air of both poultry houses ranged between 4 x 101– 7.2 x 103 for mesophilic bacteria, 0– 1.3 x 104 for staphylococci, 0 - 7 x 101 for coli group bacteria, and 2 x 101 – 1.3 x 104 for fungi (Fig. 2-5).
Outside poultry house I mesophilic bacteria were the most numerous organisms in the spring and summer and formed about 55% of the local microbial community. Less numerous staphylococci and moulds constituted about 17% and 27.5%, respectively. The concentration of
The level of outdoor air contamination was evaluated in the accordance with the Polish Norm (Table 1). Contaminations by mesophilic bacteria and moulds in areas surrounding poultry house I in the summer was the highest in sampling point I 10. In relation to the Polish Norms this site was heavily polluted. On the other hand, the number of staphylococci at all sampling sites in both poultry houses (except sampling points I 50 and II 100 in the spring) indicated a high contamination. In contrast, the air in the surrounding areas could be classified as medium-contaminated at all sites around the poultry houses II in the summer and in sampling point II 50 in the spring, with mesophilic bacteria. The evaluation based on the Polish Norm revealed that none of the researched measuring sites around the poultry house II was significantly contaminated by fungal microflora.
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Mesophilic bacteria |
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Fungi |
not pollution * | < 1x103 | 0 | 3x103-5x103 |
medium pollution ** | 1x103- 3x103 | <25 | 5x103-1x104 |
heavily pollution *** | "/3x103 | "/25 | "/ 1x104 |
Fifteen species of bacteria detected in the indoor air represented 10 genera
In this work we detected 30 species (16 in poultry houses and 23 in surrounding areas) representing 16 fungal genera:
Among fungi identified from the farm I, there distinctly dominated the species belonged to genera
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4. Discussion
The literature data usually show the air biopollutant concentration inside the poultry houses. According to many studies (Agranovski et al., 2007, Radon et al., 2002, Vučemilo et al., 2006, 2007) the number of bacteria in poultry houses ranged from 103 to 1010 CFU/m3, and the concentrations of fungi was from 2.5 x 101 to 4.9 x 106 CFU/m3. Much less is known about the relationships between the indoor and the outdoor biological pollution. In comparison with the rate of microorganisms contamination in the poultry houses, the concentration of bacterial and fungi in the air in surrounding areas was considerable lower and did not exceed the values found inside farms. Airborne bacterial and fungi levels measured in poultry houses I and II were always higher than in adjacent areas. Therefore we can suggested that the source of microorganisms are probably the farm objects. Baykov & Stoyanov 1999 also reported higher bacterial levels inside broiler farms than in nearby areas and the average values were similar to our results, i.e. value of mean of 16020 CFU/m3 for farmhouses and range of2060 CFU/m3to 386 CFU/m3 for immediate areas (10 m and 100 m from farm object, respectively). Very high concentration of microorganisms may reflect on insufficient ventilation in relation to the number of animals kept in poultry houses.
The presence of bacteria and fungi in poultry air is a natural phenomenon. Their primary source are the animals themselves, feed and litter. Microorganisms are a constituent of solid and liquid bioaerosols. This mostly refers to saprophytes, however pathogenic microorganisms were also found in the poultry houses air. Aerial count of pathogenic bacteria and fungi depends on the health condition of animals kept in the poultry houses. In addition, microorganisms count inside poultry farms air and monitoring of its emission from this building to the adjacent environment are important parameters for the assessment of the influence of poultry houses on the environmental pollution (Matković et al., 2006). In the present study, microbiological air contamination were determined at three sites at a distance of 10 m, 50 m and 100 m from the poultry houses. The results of total microorganisms count measurements outside the both poultry houses showed it to be lower than the total bacterial and moulds count inside the poultry houses. Number of microorganisms increase at 10 m distance from the poultry houses and gradually decreased to reach the lowest value at a distance of 100 m.
Analyzing the results of the microbiological research about air pollution, it should be remembered that the results are temporary values, occurring at the moment the measurement. In connection with the physico-chemical properties of the air, the degree of contamination of the air can change diametrically within a few minutes (Donderski et al., 2005). Weather condition have a enormous influence on the count of microorganisms in the air. Temperature rise accompanied by rain scarcity can lead to a sudden increase in the concentration of microorganisms in the air. Consequently in summer with the weather conditions most friendly for the spread and development of numerous microorganisms, mesophilic bacteria, staphylococci, gram-negative bacteria and fungi were the more abundant in the air around the poultry houses than in early spring, where the temperature and humidity were lower.
Staphylococci seem to be a useful indicator bacteria (Schulz et al., 2004). Although this group of bacteria do not produce spores, they have the ability to survive in the air for a long time, which means spreading infections through the air. In the poultry houses I and II and outdoor air we observed very high numbers of potentially pathogenic staphylococci, what is really negative phenomenon. In our study the predominant species were coagulase negative.
Moulds and yeast can live practically anywhere and have particularly favorable conditions inside the poultry houses. Among fungi recovered from the farm I, the species belonged to genera
In the air surroundings poultry house II dominated species were
The highest total
5. Conclusions
The farming buildings are emitters of the considerable amounts of microbiological contaminants into the atmospheric air. This high emission of potentially pathogenic microorganisms via aerosols from animal housing facilities to the outdoor environment may constitute a considerable risk to human health and environmental pollution.
So far, in literature there are no reliable data about relationships between the indoor and the outdoor biological pollution. This study contributes to the understanding of the level concentration of bioaerosol and its composition with regard to the different distance from farms. The quantity and quality of microbial analysis shows both different bacteria genera and fungi in indoor and outdoor microbiological contamination. Comparing our own results with available literature data on the indoor and outdoor air biopollutant concentration in the poultry houses enables to understand the distribution process.
References
- 1.
Agranovski V. Reponen T. Ristovski Z. 2007 Survey of bioaerosol emissions from Australian poultry buildings. European Aerosol Conference. Salzburg, Abstract, 28. - 2.
Araujo R. Cabral J. P. 2010 Fungal air quality in medical protected environments. In: , Ashok Kumar,357 382 Sciyo,978-9-53307-131-2 Publisher: Sciyo. - 3.
Bakutis B. Monsteviliene E. Januskeviciene G. 2004 Analyses of airborne contamination with bacteria, endotoxins and dust in livestock barns and poultry houses. ,73 283 289 - 4.
Baykov B. Stoyanov M. 1999 Microbial air pollution caused by intensive broiler chicken breeding. . Microbiology ecology,29 389 392 - 5.
Crook B. Easterbrook A. Stagg S. 2008 Exposure to dust and bioaerosols in poultry farming. Summary of observations and data. Prepared by the Health and Safety Laboratory for the Health and Safety Executive. - 6.
Dehoog S. Guarro J. Gene J. Figureas M. J. 2008 Atlas of Clinical Fungi. Centraalbureall voor Schiielcultures/ Universitat Rovira i Virgili. - 7.
Donderski W. Walczak M. Pietrzak M. 2005 Microbiological contamination of fair within the city of Toruń. ,14 2 223 230 - 8.
Dutkiewicz J. 1987 Bacteria in farming environment. ,71 154 71 88 - 9.
Ejdys E. Michalak J. Szewczyk K. M. 2009 Yeast-like fungi isolated from indosr air in school buildings and the surrounding outdoor air. Acta Mycologica,44 1 97 107 - 10.
Lonc E. Plewa K. 2009 Microbiological air contamination in poultry houses,14 4 445 449 - 11.
Karwowska E. 2005 Microbiological air contamination in farming environment..,19 1 15 19 - 12.
Kwaśna H. Chełkowski J. Zajkowski P. 1991 Grzyby (Mycota). Tom XXII. PAN, Warszawa Kraków, 136 ss. - 13.
Lues J. Theron M. Venter P. Rasephei H. 2007 Microbial composition in bioaerosols of a high-throughput chicken-slaughtering facility. Poultry Science,86 142 149 - 14.
Pasqualotto A. 2008 Differences in pathogenicity and clinical syndromes due to and Aspergillus flavus. Medical mycology,1 10 - 15.
Polish Norm (PN-89/Z-04111/02). Air purity protection. Microbiological testing. Determination number of the bacteria in the atmospheric air (emission) with sampling by aspiration and sedimentation methods. - 16.
Polish Norm (PN-89/Z-04111/03). Air purity protection. Microbiological testing.Determination number of the fungi in the atmospheric air (emission) with sampling by aspiration and sedimentation methods. - 17.
Matković K. Vučemilo M. Vinković B. Pavičić Ž. Matković S. Benić M. 2009 Airborne fungi in a dairy barn with emphasis on microclimate and emission. ,79 3 207 218 - 18.
Matković K. Vučemilo M. Vinković B. Šeol B. Pavičić Ž. Matković S. Tofant A. Matković S. 2006 Effect of microclimate on bacterial count and airborne emission from dairy barns on the environment. ,13 349 354 - 19.
Nevalainen A. 2007 Bio-aerosols as exposure agents in indoor environments in relation to asthma and allergy. National Public Health Institute, Department of Environmental Health POB 95, FI-70701 Kuopio, Finland. - 20.
Pomorska D. Larsson L. Skórska C. Sitkowska J. Dutkiewicz J. 2009 Levels of bacterial endotoxins in the samples of settled dust collected in animal houses. ,53 37 41 - 21.
Radon K. Danuser B. Iversen M. Monso O. Weber Ch. Hartung J. Palmgren U. Nowak D. 2002 Air contaminants in different European farming environments,9 41 48 - 22.
Romanowska-Słomka I. Mirosławski J. 2009 Biological hazards in an industrial poultry farm- research results. ,7 16 19 - 23.
Raper K. B. Thom Ch. Fennell D. I. 1949 A manual of the Penicillia. The Williams & Wilkins Company, Baltimore, USA. - 24.
Raper K. B. Fennell D. I. 1965 The genus Aspergillus. The Williams & Wilkins Company, Baltimore, USA. - 25.
Salo P. Arbes S. Sever M. Jaramillo R. Cohn R. London S. Zeldin D. 2006 Exposure to Alternaria alternata in US homes is associated with asthma symptoms. ,118 4 892 898 - 26.
Schierl R. Heise A. Egger U. Schneider F. Eichelser F. Neser S. Nowak D. 2007 Endotoxin concentration in modern animal houses in southern Bavaria. ,14 1 129 136 - 27.
Schulz J. Hartung J. Seedorf L. Formosa C. 2004 Staphylococci as an indicator for bacterial emissions from a broiler house. ,75 78 - 28.
Soliman S. Sobeih M. Hussein M. Abdel-Latiff H. Moneim A. 2009 Seasonal epidemiological surveillance on bacterial and fungal pathogens in broiler farms in Egypt.8 8 720 727 - 29.
Siemińki M. 2001 , Wydawnicto Naukowe PWN,8-30113-561-1 - 30.
Srikanth P. Suchithra S. Steinberg R. 2008 Bio-aerosols in indoor environment: composition, health effects and analysis. Indian Journal of Medical Microbiology,26 4 302 312 - 31.
Vučemilo M. Vinković B. Matković K. 2006 Influence of broiler age on airborne pollutant content in poultry house. ,48 3 6 - 32.
Vučemilo M. Matković K. Vinković B. Jakšić S. Granić K. Mas N. 2007 The effect of animal age on air pollutant concentration in a broiler house. .52 6 170