Slaughterhouses Fungal Burden Assessment: A Contribution for the Pursuit of a Better Assessment Strategy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Assessed Settings
2.2. Sample Collection
2.2.1. Conventional Methodologies
2.2.2. Molecular Methodologies
2.3. Sample Preparation and Analysis
2.3.1. Conventional Methodologies
2.3.2. Molecular Methodologies
2.4. Data Analysis
3. Results
3.1. Fungal Load
3.2. Fungal Identification
3.2.1. Poultry Slaughterhouse
3.2.2. Swine and Bovine Slaughterhouse
3.2.3. Large Animal Slaughterhouse
3.3. Fungal Detection
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Poultry Slaughterhouse (PS) | Swine/Bovine Slaughterhouse (SBS) | Large Animal Slaughterhouse (LAS) |
---|---|---|
Birds hanging | Gut room swine | Bovine line |
Reception | Gut room bovine | Paws room |
Stacker | Bleeding swine | Heads room |
Bleeding | Cutting swine | Gut room |
Evisceration | Bleeding bovine | Expedition |
Cutting | Cutting bovine | Barn |
Slaughterhouses | Conventional Methods | Molecular Biology | Fungal Species Assessed by Molecular Biology | |
---|---|---|---|---|
Air Samples | Surface Samples | Air Samples | ||
Poultry | 6 | 6 (floor) | 6 | A. flavus complex (toxigenic strains) A. fumigatus complex A. ochraceus complex |
Swine | 6 | 6 (walls) | 6 | |
Large animal | 6 | 6 (floor) | 5 | |
Total of samples | 18 | 18 | 17 |
Fungal Species Targeted | Sequences |
---|---|
A. flavus complex (Toxigenic Strains) | |
Primer Forward | 5‘-GTCCAAGCAACAGGCCAAGT-3‘ |
Primer Reverse | 5‘-TCGTGCATGTTGGTGATGGT-3‘ |
Probe | 5‘-TGTCTTGATCGGCGCCCG-3‘ |
A. fumigatus complex | |
Primer Forward | 5‘-CGCGTCCGGTCCTCG-3‘ |
Primer Reverse | 5‘-CGTGAGGCGGGAGCA-3‘ |
Probe | 5‘-CCAACCTCCCACCCGTG-3‘ |
A. ochraceus complex | |
Primer Forward | 5‘-CGGGTCTAATGCAGCTCCAA-3‘ |
Primer Reverse | 5‘-CGGGCACCAATCCTTTCA-3‘ |
Probe | 5‘-CGTCAATAAGCGCTTTT-3‘ |
Fungal Identification | Fungal Quantification |
---|---|
Indoor air samples (PS) | (CFU/m3; %) |
Scopulariopsis sp. | 950; 59.52 |
Penicillium sp. | 524; 32.83 |
Aspergillus fumigatus complex | 50; 3.13 |
Others | 72; 4.5 |
Indoor surfaces samples (PS) | (CFU/m2; %) |
Mucor sp. | 10,000; 100 |
Indoor air samples (SBS) | (CFU/m3; %) |
Cladosporium sp. | 370; 45.7 |
Penicillium sp. | 270; 33.3 |
Aureobasidium sp. | 90; 11.1 |
Others | 80; 9.9 |
Indoor air samples (LAS) | (CFU/m3; %) |
Penicillium sp. | 118; 80.8 |
Aspergillus ochraceus complex | 8; 5.5 |
Cladosporium sp. | 8; 5.5 |
Others | 12; 8.2 |
Indoor surfaces samples (LAS) | (CFU/m2; %) |
Scopulariopsis brumptii | 40,000; 40 |
A. terreus complex | 30,000; 30 |
Others | 30,000; 30 |
Sampling Sites | Fungal Quantification | Fungal Detection | |
---|---|---|---|
Poultry slaughterhouse (PS) | Air (CFU/m3) | Surfaces (CFU/m2) | Real-time PCR (Ct, cycle threshold) |
Birds hanging | - | - | n. d. |
Reception | - | - | n. d. |
Stacker | 10 | - | 35.7 |
Bleeding | 30 | - | 35.92 |
Evisceration | 10 | - | 36.53 |
Cutting | - | - | n.d. |
Swine and bovine slaughterhouse (SBS) | Air (CFU/m3) | Surfaces (CFU/m2) | Real-time PCR (Ct, cycle threshold) |
Gut room swine | - | - | 33.91 |
Gut room bovine | 10 | - | n.d. |
Bleeding swine | - | - | 33.53 |
Cutting swine | - | - | 33.61 |
Bleeding bovine | - | - | n.d. |
Cutting bovine | 35.89 | ||
Large animal slaughterhouse (LAS) | Air (CFU/m3) | Surfaces (CFU/m2) | Real-time PCR (Ct, cycle threshold) |
Bovine line | - | - | n.d. |
Paws room | - | - | 35.85 |
Heads room | - | - | n. d. |
Gut room | - | - | n.d. |
Expedition | - | - | 34.47 |
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Viegas, C.; Faria, T.; Dos Santos, M.; Carolino, E.; Sabino, R.; Quintal Gomes, A.; Viegas, S. Slaughterhouses Fungal Burden Assessment: A Contribution for the Pursuit of a Better Assessment Strategy. Int. J. Environ. Res. Public Health 2016, 13, 297. https://doi.org/10.3390/ijerph13030297
Viegas C, Faria T, Dos Santos M, Carolino E, Sabino R, Quintal Gomes A, Viegas S. Slaughterhouses Fungal Burden Assessment: A Contribution for the Pursuit of a Better Assessment Strategy. International Journal of Environmental Research and Public Health. 2016; 13(3):297. https://doi.org/10.3390/ijerph13030297
Chicago/Turabian StyleViegas, Carla, Tiago Faria, Mateus Dos Santos, Elisabete Carolino, Raquel Sabino, Anita Quintal Gomes, and Susana Viegas. 2016. "Slaughterhouses Fungal Burden Assessment: A Contribution for the Pursuit of a Better Assessment Strategy" International Journal of Environmental Research and Public Health 13, no. 3: 297. https://doi.org/10.3390/ijerph13030297