The association between faecal host DNA or faecal calprotectin and feed efficiency in pigs fed yeast-enriched protein concentrate
Gut cell losses contribute to overall feed efficiency due to the energy requirement for cell replenishment. Intestinal epithelial cells are sloughed into the intestinal lumen as digesta passes through the gastrointestinal tract, where cells are degraded by endonucleases. This leads to fragmented DNA being present in faeces, which may be an indicator of gut cell loss. Therefore, measuring host faecal DNA content could have potential as a non-invasive marker of gut cell loss and result in a novel technique for the assessment of how different feed ingredients impact upon gut health. Faecal calprotectin (CALP) is a marker of intestinal inflammation.
This was a pilot study designed to test a methodology for extracting and quantifying DNA from pig faeces, and to assess whether any differences in host faecal DNA and CALP could be detected. An additional aim was to determine whether any differences in the above measures were related to the pig performance response to dietary yeast-enriched protein concentrate (YPC). Newly weaned (∼26.5 days of age) Large White × Landrace × Pietrain piglets (8.37 kg ±1.10, n = 180) were assigned to one of four treatment groups (nine replicates of five pigs), differing in dietary YPC content: 0% (control), 2.5%, 5% and 7.5% (w/w). Pooled faecal samples were collected on days 14 and 28 of the 36-day trial. Deoxyribonucleic acid was extracted and quantitative PCR was used to assess DNA composition. Pig genomic DNA was detected using primers specific for the pig cytochrome b (CYTB) gene, and bacterial DNA was detected using universal 16S primers. A pig CALP ELISA was used to assess gut inflammation.
Dietary YPC significantly reduced feed conversion ratio (FCR) from weaning to day 14 (P<0.001), but not from day 14 to day 28 (P = 0.220). Pig faecal CYTB DNA content was significantly (P = 0.008) reduced in YPC-treated pigs, with no effect of time, whereas total faecal bacterial DNA content was unaffected by diet or time (P>0.05). Faecal CALP levels were significantly higher at day 14 compared with day 28, but there was no effect of YPC inclusion and no relationship with FCR. In conclusion, YPC reduced faecal CYTB DNA content and this correlated positively with FCR, but was unrelated to gut inflammation, suggesting that it could be a non-invasive marker of gut cell loss. However, further validation experiments by an independent method are required to verify the origin of pig faecal CYTB DNA as being from sloughed intestinal epithelial cells.
Human Stromal Cell Aggregates Concentrate Adipose Tissue Constitutive Cell Population by In Vitro DNA Quantification Analysis
Background: Regenerative cell strategies rely on stromal cell implants to attain an observable clinical outcome. However, the effective cell dose to ensure a therapeutic response remains unknown. To achieve a higher cell dose, the authors hypothesized that reducing the volume occupied by mature adipocytes in lipoaspirate will concentrate the stromal vascular fraction present in the original tissue.
Methods: Human standardized lipoaspirate (n = 6) was centrifuged (1200 g for 3 minutes) and the water phase was discarded. Mechanical disaggregation was achieved by shearing tissue through 2.4- and 1.2-mm Luer-to-Luer transfers. After a second centrifugation (800 g for 10 minutes), stromal cell aggregates were separated from the supernatant oil phase. Lipoaspirate percentage composition was determined by its constituent weights. Cell content was measured by total DNA quantification, and partial cell viability was determined by image cytometry. Tissue sections were evaluated histologically (hematoxylin and eosin and Masson trichrome stains).
Results: Stromal cell aggregates reduced the standardized lipoaspirate mass to 28.6 ± 4.2 percent. Accordingly, the cell density increased by 222.6 ± 63.3 percent (from 9.9 ± 1.4 million cells/g to 31.3 ± 6.6 million cells/g; p < 0.05). Cell viability was unaffected in stromal cell aggregates (71.3 ± 2.5 percent) compared to standardized lipoaspirate (72.2 ± 2.3 percent), and histologic analysis revealed high-density areas enriched with stromal cells (622.9 ± 145.6 percent) and extracellular matrix (871.2 ± 80.3 percent).
Conclusion: Stromal cell aggregates represent a biological agent that triplicates the cell density versus unprocessed lipoaspirate, low on oil and water fluids, and enriched extracellular matrix components.
Human Stromal Cell Aggregates Concentrate Adipose Tissue Constitutive Cell Population by In-Vitro DNA Quantification Analysis
Background: Regenerative cell strategies rely on stromal cell implants to attain an observable clinical outcome. Yet, the effective cell dose to ensure a therapeutic response remains unknown. To achieve a higher cell dose, we hypothesized that reducing the volume occupied by mature adipocytes in lipoaspirate will concentrate the stromal vascular fraction present in the original tissue.
Methods: Human standardized lipoaspirate (n = 6) was centrifuged (1200 g for 3 min) and the water phase was discarded. Mechanical disaggregation was achieved by shearing tissue through 2.4 and 1.2 mm Luer-to-Luer transfers. After a second centrifugation (800 g for 10 min), stromal cell aggregates were separated from the supernatant oil phase. Lipoaspirate percentage composition was determined by its constituent weights. Cell content was measured by total DNA quantification and partial cell viability was determined by image cytometry. Tissue sections were evaluated histologically (hematoxylin-eosin and Masson’s trichrome).
Results: Stromal cell aggregates reduced the standardized lipoaspirate mass to 28.6 ± 4.2 percent. Accordingly, the cell density increased by 222.6 ± 63.3 percent (from 9.9 ± 1.4 to 31.3 ± 6.6 million cells/g, p < 0.05). Cell viability was unaffected in stromal cell aggregates (71.3 ± 2.5 percent) compared to standardized lipoaspirate (72.2 ± 2.3 percent) and histologic analysis revealed large density areas enriched with stromal cells (622.9 ± 145.6 percent) and extracellular matrix (871.2 ± 80.3 percent).
Conclusions: Stromal cell aggregates represent a biological agent that triplicates the cell density versus unprocessed lipoaspirate, low on oil and water fluids, and enriched extracellular matrix components
Distribution and degradation of DNA from non-genetically and genetically modified soybean (Roundup Ready): Impact of soybean protein concentrate and soybean protein isolate preparation
To improve genetically modified product labelling legislation and promote the development of genetically modified foods, the mass variations of genomic DNA and length distributions of DNA fragments in non-genetically and genetically modified soybean (Roundup Ready) and the variations in transgenic contents during soybean protein concentrate (SPC) and soybean protein isolate (SPI) preparation were monitored. The material masses throughout the process conformed to the law of mass conservation, and amounts of DNA recovered decreased with SPC and SPI preparation.
The successive steps of ethanol extraction led to a decrease in the size distribution of the recovered DNA. For the LECTIN and CP4 EPSPS targets investigated, longer fragments (>800 bp) were more affected than smaller fragments (<200 bp), and both targets degraded slowly upon degradation into small fragments. DNA distribution and degradation thereby affected GMO quantification. After preparation procedures, the transgenic contents of SPC and SPI products were higher than that of raw soybean.
CHO DNA Concentrate |
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D552 | Cygnus Technologies | 1 ml | 650.4 EUR |
CHO DNA Concentrate |
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D556 | Cygnus Technologies | 120 ul | 562.8 EUR |
E.coli DNA Concentrate |
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D412 | Cygnus Technologies | 1 ml | 650.4 EUR |
Human DNA Concentrate |
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D162 | Cygnus Technologies | 1 ml | 607.2 EUR |
NS/0 DNA Concentrate |
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D222 | Cygnus Technologies | 1 ml | 650.4 EUR |
Liver Concentrate |
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L04640 | Pfaltz & Bauer | 500G | 179.5 EUR |
NADPH Concentrate, 300UL |
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X044-300UL | Arbor Assays | 300UL | 59 EUR |
NADPH Concentrate, 500UL |
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X044-500UL | Arbor Assays | 500UL | 73 EUR |
PBS (10X Concentrate) |
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85R-125 | Fitzgerald | 250 ml | 199 EUR |
PBS (10X Concentrate) |
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MBS539639-250mL | MyBiosource | 250mL | 345 EUR |
PBS (10X Concentrate) |
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MBS539639-5x250mL | MyBiosource | 5x250mL | 1400 EUR |
NADH Concentrate, 1.2ML |
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X090-1.2ML | Arbor Assays | 1.2ML | 59 EUR |
Cholesterol Concentrate |
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MBS173582-10g | MyBiosource | 10g | 1275 EUR |
Cholesterol Concentrate |
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Cholesterol Concentrate |
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Cholesterol Concentrate |
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MBS173645-INQUIRE | MyBiosource | INQUIRE | Ask for price |
NADPH Concentrate, 1ML |
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X043-1ML | Arbor Assays | 1ML | 85 EUR |
NADPH Concentrate, 1.4ML |
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X044-1.4ML | Arbor Assays | 1.4ML | 109 EUR |
Ceraplex Lipid Concentrate(1000X) |
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C3316-010 | GenDepot | 100ml | 144 EUR |
Ceraplex Lipid Concentrate(1000X) |
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C3316-050 | GenDepot | 500ml | 295.2 EUR |
AMP Buffer Concentrate |
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40120060-1 | Glycomatrix | 1 L | 66.75 EUR |
AMP Buffer Concentrate |
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40120060-2 | Glycomatrix | 4 L | 223.5 EUR |
AMP Buffer Concentrate |
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40120060-3 | Glycomatrix | 500 mL | 21.93 EUR |
AMP buffer concentrate |
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ADB0030 | Bio Basic | 500ml | 79.84 EUR |
Wash Buffer Concentrate, 125ML |
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X007-125ML | Arbor Assays | 125ML | 168 EUR |
BSA Antigen Concentrate |
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F031G | Cygnus Technologies | 1 ml | 262.8 EUR |
The association between faecal host DNA or faecal calprotectin and feed efficiency in pigs fed yeast-enriched protein concentrate.
- Gut cell losses contribute to overall feed efficiency due to the energy requirement for cell replenishment. Intestinal epithelial cells are sloughed into the intestinal lumen as digesta passes through the gastrointestinal tract, where cells are degraded by endonucleases. This leads to fragmented DNA being present in faeces, which may be an indicator of gut cell loss. Therefore, measuring host faecal DNA content could have potential as a non-invasive marker of gut cell loss and result in a novel technique for the assessment of how different feed ingredients impact upon gut health. Faecal calprotectin (CALP) is a marker of intestinal inflammation.
- This was a pilot study designed to test a methodology for extracting and quantifying DNA from pig faeces, and to assess whether any differences in host faecal DNA and CALP could be detected. An additional aim was to determine whether any differences in the above measures were related to the pig performance response to dietary yeast-enriched protein concentrate (YPC). Newly weaned (∼26.5 days of age) Large White × Landrace × Pietrain piglets (8.37 kg ±1.10, n = 180) were assigned to one of four treatment groups (nine replicates of five pigs), differing in dietary YPC content: 0% (control), 2.5%, 5% and 7.5% (w/w). Pooled faecal samples were collected on days 14 and 28 of the 36-day trial.
- Deoxyribonucleic acid was extracted and quantitative PCR was used to assess DNA composition. Pig genomic DNA was detected using primers specific for the pig cytochrome b ( CYTB ) gene, and bacterial DNA was detected using universal 16S primers. A pig CALP ELISA was used to assess gut inflammation. Dietary YPC significantly reduced feed conversion ratio (FCR) from weaning to day 14 (P0.05). Faecal CALP levels were significantly higher at day 14 compared with day 28, but there was no effect of YPC inclusion and no relationship with FCR. In conclusion, YPC reduced faecal CYTB DNA content and this correlated positively with FCR, but was unrelated to gut inflammation, suggesting that it could be a non-invasive marker of gut cell loss. However, further validation experiments by an independent method are required to verify the origin of pig faecal CYTB DNA as being from sloughed intestinal epithelial cells.