Proceedings available at: http://www.extension.org/67695 Manure generated from concentrated animal feeding operations may serve as a source of steroids in surface water and adversely impact the development of aquatic ecosystems. The objectives of this research were to determine the amount of steroids and metabolites in manure from beef cattle production pens, and runoff from crop production fields.

1. Waste of Worth
April, 2013
Denver, CO
Charles A. Shapiro, Sagor Biswas, David D. Tarkalson, William L.
Kranz, David P. Shelton, Terry L. Mader, Daniel D. Snow, Simon J. van
Donk, Shannon L. Bartelt-Hunt, and Tian C. Zhang
Technical assistance: Leslie Johnson and Mike Mainz
University of Nebraska-Lincoln
Email: cshapiro@unl.edu

2. Funding provided by US-EPA -2007program
Beef and dairy
Poultry
litter
Poultry
broiler
Beef, dairy, poultry and swine

3.  96% of CAFO cattle
use implants
 CONCERN: Potential for
natural and synthetic
steroid hormones in
livestock waste to reach
groundwater and
surface waters
?

4.  Endogenous (natural)
 Estrodiol
 Estrone
 Testosterone
 Androstenedione
 Progesterone
 Exogenous (synthetic)
 Trenbolone
 Zearalonol
 Melengestrol Acetate
17 -estradiol
estrone
Testosterone Progesterone
-zearalonol
Melengestrol acetate17 -trenbolone

5. RalgroTM Implant active ingredient

6.  Increase average daily gain (ADG)
 5 - 25 %
 Improve feed efficiency (G/F)
 5 - 18 %
 Improve lean tissue deposition
 5 - 12 %

7. 1) Quantify hormones in various stages of the manure
pathway in cattle feedlots.
2) Determine the effects of different handling practices of
cattle feedlot wastes on the stability and availability of
hormones.
3) Determine the effects of different land application
strategies on the fate and transport of hormones in runoff
and vadose zone soils.
4) Determine if grasses grown in conservation buffers
assimilate hormones.

8. 8
Meat
Market
Hormone supplements:
36 mg α-zearalanol,
140 mg of TBA and 14 mg
of 17β-estradiol (after 35 days)
0.45 mg of MGA (daily feed, 145 days)
Fresh Manure ~25kg/day
~0.5 mg/day excretion
~ 20 ug/kg maximum
Compost Stockpile
Runoff Holding
Pond
Land
Application
Overland
runoff
Soil Water
Movement
Plant Uptake
Streams, Lak
es
Vadose zone,
Ground water
Animal
Ingestion

9.  96 heifers split between six pens
 Half receive implants /feed
supplement
 Surface samples, feces and urine
spots collected and analyzed
(7, 45, 125 days)
 Feed lot run-off from natural rainfall
events collected using tipping bucket
sampler
 Pens scraped and waste stockpiled
or composted
 Feeding pen study conducted 2007
and 2008

10.  Low concentrations of steroid hormones
detected in feeding pen run-off
 Steroids detected in ≥ 80% of treated and control
samples
 Synthetic steroids only detected in runoff from
treated groups
 Mostly endogenous steroids and possibly mycotoxins
detected in dissolved fraction
 Large proportion held in feed-lot surfaces
 Seasonal variation observed in metabolites and
concentrations

11. 11
Meat
Market
Livestock
(Heifers)
Hormone Supplements:
36 mg α-zearalanol,
140 mg of TBA and 14 mg of 17β-
estradiol (after 35 days)
0.45 mg of MGA (daily feed)
Manure
Compost Stockpile
Runoff Holding
Pond
Land
Application
Overland
runoff
Soil Water
Movement
Plant Uptake
Streams, Lak
es
Vadose
zone, Ground
water
Animal
Ingestion

12.  Feedlot study initiated June
 Waste handling study initiated October
 Manure from
treated and
untreated cattle
placed in
compost piles
and anaerobic
stockpiles
 Piles turned
through Spring

13. Androgens Mycotoxins Estrogens Progestagens
Manure
4-androstenedione
Androsterone
α-zearalenol
β-zearalenol
α-zearalanol
17α-estradiol
17β-estradiol
Estriol
Estrone
Melengesterol
acetate
Progesterone
17α-
hydroxyprogesterone
TotalConcentration
ng g-1
CU 1.8 ND ND ND ND ND ND 2.0 5.1 <0.5 0.9 ND 9.8
CT 3.3 3.5 ND ND 1.0 <0.5 <0.5 ND 5.4 <0.5 0.7 <0.5 13.9
SU 6.8 ND ND 8.8 0.8 ND <0.5 ND 4.3 ND 4.0 ND 24.7
ST 5.3 ND 1.9 11.4 ND <0.5 ND ND 3.7 <0.5 4.1 <0.5 26.4
13
Detection limit: 0.5 ng g-1
CU= Compost untreated, CT= Compost treated, SU= Stockpile untreated, ST=
Stockpile treated, ND= Non-detectable
Average steroid concentration in manure sample from April 2008

14.  To evaluate the effects of:
 precipitation (simulation at 1 DAT and 30 DAT)
 manure handling (composting vs stockpiling,
treated vs untreated)
 tillage (disk, plow and no-till)
-on the presence of steroid hormones in overland
runoff
14

15.  Study area: Haskell
Agricultural Laboratory,
Concord
 Soil: Nora silty clay loam
(28% sand, 48% silt and
24% clay)
 Slope: 8%
 Average annual
precipitation: 672 mm
(26 in)
 Average annual
temperature: 8 C (47 F)
15
Feedlot

16.  Manure application rate:
 193 Mg ha-1 stockpiled
 159 Mg ha-1 composted
 N availability factor: 0.15
 N requirement: 170 kg-N
ha-1 for dryland corn
 Rainfall intensity:
75 mm hr-1
 6 runoff samples
collected at 5 mins
interval after runoff
initiation
16
Experimental Design

17. 17

18. 18
Results (2008):
b b b
c
b
a
0
30
60
90
120
1 DAT 30 DAT 1 DAT 30 DAT 1 DAT 30 DAT
Timetorunoffinitiation(min)
(A) Tillage Treatment

19. 19
Results (2008):
ab
a
ab
b
0
20
40
60
80
100
120
1 DAT 30 DAT 1 DAT 30 DAT
Timetorunoffinitiation
(min)
(B) Manure Treatment

20. 20
Results:
a
b
a
a
a
b
0
4
8
12
16
1 DAT 30 DAT 1 DAT 30 DAT 1 DAT 30 DAT
Runoff(mm)
(A) Tillage Treatment

21. 0
5
10
15
20
25
30
Disk Plow No-till
Frequencyofdetection(%)
1 DAT 30 DAT
0
5
10
15
20
25
30
Compost Stockpile Treated UntreatedFrequencyofdetection(%)
1 DAT 30 DAT

22.  Lack of detection in the 2008
 Ensure sufficient concentrations to
detect effect of treatments
 Not used or released by beef cattle
 17α-Ethynylestradiol (EE2)
22

23.  Manure application
 193 Mg ha-1
 N availability factor:
0.15
 170 kg-N ha-1 N
requirements
 Application rate of 7.5
mg m-2
 Using a backpack sprayer
with a 3 m boom
 RCB with three
replications 23

24. 24
Runoff
initiation
time (min)
Total
Precipitation
(mm)
Pr>F
Treatment
Tillage 0.27 0.10
Manure 0.19 0.24
EE2 0.86 0.81
Mean
NT 53 100
T 81 133

25. Flow Weighted
Average Concentration
(ng L-1)
Mass
transport
(mg ha-1)
Treatment Pr>F
Tillage 0.0003 0.003
Manure 0.05 0.11
Tillage*Manure 0.83 0.90
Mean
NT-EE2 with Manure 1765 a 110 a
T-EE2 with Manure 62 b 3 b

26.  Little steroid remains in
manure after feedlot
 Minor benefit from
composting
 Runoff from fields affected by
tillage and timing
 Interaction between time from
tillage to rainfall affects erosion
and hence,
 Size of storm to produce similar
potential losses varies
 Little leaching, held in soil
(data not shown)
Livestock
(Heifers)
Manure
Compost Stockpile
Land
Applicati
on
Overland
runoff
Streams,
Lakes