inhibition of total gas production, methane, hydrogen sulfide, and sulfate-reducing bacteria from in...
DESCRIPTION
For more: http://www.extension.org/67635 Management practices from large-scale swine production facilities have resulted in the increased collection and storage of manure for off-season fertilization use. Odor produced during storage has increased the tension among rural neighbors and among urban and rural residents, and greenhouse gas emissions may contribute to climate change. Production of these compounds from stored manure is the result of microbial activity of the anaerobic bacterial populations present during storage. We have been studying the bacterial populations of stored manure to develop methods to reduce bacterial metabolic activity and production of gaseous emissions, including the toxic odorant hydrogen sulfide produced by sulfate-reducing bacteria. Quebracho and other condensed tannins were tested for effects on total gas, hydrogen sulfide, and methane production and levels of sulfate-reducing bacteria in in vitro swine manure slurries. Quebracho condensed tannins were found to be most effective of tannins tested, and total gas, hydrogen sulfide, and methane production were all inhibited by greater than 90% from in vitro manure slurries. The inhibition was maintained for at least 28 days. Total bacterial numbers in the manure were reduced significantly following addition of quebracho tannins, as were sulfate-reducing bacteria. These results indicate that the condensed tannins are eliciting a collective effect on the bacterial population, and the addition of quebracho tannins to stored swine manure may reduce odorous and greenhouse gas emissions.TRANSCRIPT
USDA - ARS - National Center for Agricultural Utilization Research
INHIBITION OF TOTAL GAS INHIBITION OF TOTAL GAS PRODUCTION, METHANE, HYDROGEN PRODUCTION, METHANE, HYDROGEN
SULFIDE, AND SULFATE-REDUCING SULFIDE, AND SULFATE-REDUCING BACTERIA FROM IN VITRO STORED BACTERIA FROM IN VITRO STORED
SWINE MANURE USING CONDENSED SWINE MANURE USING CONDENSED TANNINSTANNINS
Terence R. WhiteheadTerence R. Whitehead
Bioenergy Research UnitBioenergy Research Unit
USDA-ARS-NCAUR, Peoria, IllinoisUSDA-ARS-NCAUR, Peoria, Illinois
Pig House
Manure Storage
Underground Storage ofSwine Manure
Partial List of Identified Volatile Organic Compounds From
Swine Manure
1-Methylethanol Acetic AcidPropanol Propanoic acid2-Methoxyethanol 2-Methylpropanoic acid2-Methylpropanol Butanoic acidButanol Hexanoic acid3-Methylbutanol 2-PhenylethanolPentanol p-Cresol3-OH-butanone 3-Methylindole3-Octanol p-Ethylphenol2,3,4-Trithiapentane o-Methoxyphenol
Additional Emissions Also Which May Play Important Roles From Swine
Manure
• Ammonia
•Hydrogen Sulfide
•Methane
Identification of Bacteria Involved inManure Breakdown and Odor Production
I. Classical Microbiology
• Pure culture isolation and identification using habitat simulating media, biochemical techniques and 16S rDNA gene sequence analyses.
II. Molecular Ecology (Culture Independent)
• Isolate total DNA from fecal and manure samples, determine DNA sequence of isolated 16S rDNA genes, and identify bacteria by comparison with known sequences in GenBank database.
Miscellaneous Parameters for Swine Manure Pits
•Temperature: 24°C
• pH: 6.85
•3-Ft. Pit: 1 x 1010 Cells/ml
• Approx.Volume: 500,000 Gallons
•Approx. Bacterial Number: 2 x 1019
Peoria Illinois USA World Manure Pit103
106
109
1012
1015
1018
1021
Comparison of Human Populations Versus Swine Manure Pit Bacteria
Summary
•Microbial populations of stored swine manure are dominated by Gram-positive, low %G+C, anaerobic bacteria.
•The vast majority of the bacterial isolates (and DNA clones) could only be identified to the genera level, and many strains represent new genera and species.
•Swine manure represents a rich ecosystem for isolation of new and novel bacteria.
• Using 16S rDNA sequence analyses and biochemical studies we have been able to identify new bacterial genera and species.
• Characterized populations of sulfate-reducing bacteria (SRB), which produce hydrogen sulfide, using PCR and qRT-PCR. Archeal 16S rDNA gene and dissimilatory sulfite reductase (dsrA) gene primer sets were used for analyses.
• Identified 3 new groups of SRB.
• Groups fell within known lineages of SRBs; however, the clones represented novel species within these genera.
Schematic representation of a condensed tannin molecule. Condensed tannins can be linear (with 4→8 bounds) or branched (with 4→6 bounds - dotted line).
In Vitro Manure Assay Bottles.
Table 1. Effects of Addition of Various Condensed Tannins on Total Gas Production by In Vitro Swine Manure Slurries Tannin % Control (Cumulative Gas Production)1
Control (No Tannin) 100.0Quebracho (0.5% w/v) 2.2C-KPN (0.5% w/v) 53.2C-KPS (0.5% w/v) 55.6NA-1 (0.5% w/v) 188.1TARA (0.5% w/v) 155.6WE-ME (0.5% w/v) 43.0_______________________________________________________1Quebracho tannin experiment terminated after 42 days. Othertannins ended after 58 days.
Inhibition of H2S Production by 0.5% Quebracho Tannins
Effects of 0.5% Quebracho Tannins on Total SRB Levels
1.0E+09
1.0E+10
1.0E+11
1.0E+12
0 1 3 7 14 21 28
Time (days)
cop
ies
16S
rD
NA
/ m
l sl
urr
ycontrol
0.5% tannins
**
**
***
Effects of 0.5% Quebracho Tannins on Bacterial Numbers
Inhibition of Methane Production by 0.5% Quebracho Tannins
Future Work: Known Knowns, Known Unknowns, and Unknown Unknowns
• Onsite Farm Testing with Quebracho Tannins
• Other Tannins Useful?
•Other Compounds for Testing? -Ionophores?
POSTER SESSION:
Combination of Borax and Quebracho Condensed Tannins Treatment to Reduce Hydrogen Sulfide, Ammonia and Greenhouse Gas Emissions from Stored Swine Manure. Yokoyama et al.
Acknowledgments
NCAUR - Peoria
Dr. Michael CottaRhonda ZeltwangerDr. Cheryl Spence
ARS – Bowling Green, KY
Dr. Kim Cook
