1. Fate of CECs in Animal Manure Amended Soils‐
Impact of Manure Application Technologies
K. Xia1*, R. O. Maguire1, and K. F. Knowlton2
1Dept. Crop and Soil Environmental Sciences, Virginia Tech.
2Dept. Dairy Sciences, Virginia Tech.
1

2.  Current manure land application technologies are
designed for N and P pollution reduction
Manure land
application methods
Broadcast
(surface application)
advantages disadvantages
Soil incorporation
tillage ‐soil compaction
‐surface crusting
& sealing
↑ carbon loss
easy operation
low cost
‐runoff
‐volatilization
low cost
↓ short term runoff
↓ volatilization (possibly)
no‐till/reduced tillage ‐specialized
equipment
‐higher cost
‐slower
↓ runoff
↓ greenhouse gas
↓ soil disturbance
↑ N, P, water use efficiency
2
‐subsurface injection

3. • Manure application method?
• Manure application timing?
• Spatial variation?
• Temporal change?
Major research questions on fate of CECs

4.  Methods and Materials
• Field‐scale rainfall simulation experiment
Study site: McCoy, Virginia
Soil type: Braddock Loam
9‐11% slope
CECs investigated:
‐ 11 hormones
‐ 4 antibiotics
4
Manure (wet weight):
‐ Dairy/cattle manure
‐ Poultry litter
56 Mg/Ha
6.7 Mg/Ha

5. 5
a)
Slits pre‐cutting

6. Treatments: Study design:
randomized complete block
3 replicates/treatmentsurface application
subsurface injection
Simulated rainfall intensity: 70 mm hr‐1
Runoff collected/plot: 30 min of runoff
Soil sampling: 0‐5 and 5‐20 cm
0, 5, 14, 60, 180 days after simulated rainfall
control without manure
6
Simulated rainfall events:
Manure
application
0 3days 7days
R2R1
R1 R1 R1
2 rain events (scheme #1, fall 2014)
1 rain event (scheme #2, fall 2015)

7.  runoff water: solid phase extraction (SPE)
• Sample extraction and cleanup
• UPLC/MS/MS analysis
 runoff sediment & soil: liquid/solid extraction
• Statistical analysis:
Tukey multiple comparison analysis in SAS
(P < 0.5 indicates significant differences) 7
11 Hormones Investigated
Hormone Class Hormones
Estrogens Estrone (E1)
17α‐estradiol (α‐E2)
17β‐estradiol (β‐E2)
Estriol (E3)
Androgens 1,4‐Androstadiene‐3,17‐dione (1‐4‐
ADD)
4‐Androstene‐3,17‐dione. (4‐AD)
Progestagens Progesterone (P)
Estrogen‐
Conjugates
Estrone‐3‐Sulfate (E1‐3S)
Estrone‐3‐Glucorinide (E1‐3G)
Estradiol‐3‐Sulfate (E2‐3S)
Estradiol‐3‐Glucorinide (E2‐3G)
4 Antibiotics Investigated
Class Compound
Lincosamide Pirlimycin
Sulfonamide Sulfamerazine
Macrolide Tylosin
Tetracycline Chlortetracycline

8.  Results
8
• Manure application method?
• Manure application timing?
• Spatial variation?
• Temporal change?

9. Concentration(ng/g,dryweightbasis)
0
50
100
150
200
250
300
350
400
Pirlimycin
Sulfamerazine
Tylosin
Chlortetracycline
Col 2
Before rain
0-5 cm
Col 2
Surface Slit 5 15 25 40
0
50
100
150
200
250
300
350
5-20 cm
distance from the slit (cm)
Subsurface injection
Surface
application
Manure application
Concentration(ng/g,d.w.)
9
• Manure application method?
CECs in soil
Surface broadcast
Manure
Soil
aggregate
Subsurface injection
Manure
Soil
aggregate
Manure aggregate
Soil aggregate
Soil organic matter
Soil microflora
CECs

10. Mass balance of pirlimycin applied with manure
water: 2.5%
sediment: 0.06%
1st rainfall (day 0)
water: 1.1%
sediment: 0.01%47%
35%
11%
7.7%
runoff
40%
56%
runoff
water: 0.05%
sediment: 0.01%
2nd rainfall (day 7)
water: 0.01%
sediment: 0%30%
9%
4%
0%
runoff
3%
17%
runoff
10
• Manure application method & impact of 2 consecutive rain events?

11. Concentration(ng/g,dryweightbasis)
0
50
100
150
200
250
300
350
400
Pirlimycin
Sulfamerazine
Tylosin
Chlortetracycline
Days between manure application rain - before rain
0 day
0-5 cm
Surface
Slit
5
15
25
40
0-5 cm
5-20 cm
5 15 25 40
0.0
0.1
0.2
5.0
10.0
5 15 25 40
0.0
0.5
1.0
Surface
Slit
5
15
25
40
0
50
100
150
200
250
300
350
5-20 cm
Surface
Slit
5
15
25
40
3 days 7 days
5 15 25 40
0.0
0.1
0.2
5.0
10.0
11
• Manure application method & timing relative to rain events?
distance from the slit (cm)
Subsurface injection
Surface
application
Surface
application
Surface
application
distance from the slit (cm)
Subsurface injection
distance from the slit (cm)
Subsurface injection
CECs in soil
Days between manure application & rain event – before rain
Concentration(ng/g,d.w.)

12. Concentration(ng/g,dryweightbasis)
0
50
100
150
200
250
300
350
400
Pirlimycin
Sulfamerazine
Tylosin
Chlortetracycline
Days between manure application rain - after rain
0 day
0-5 cm
Surface
Slit
5
15
25
40
0-5 cm
5-20 cm
5 15 25 40
0.0
0.1
0.2
5.0
10.0
5 15 25 40
0.0
0.5
1.0
Surface
Slit
5
15
25
40
0
50
100
150
200
250
300
350
5-20 cm
Surface
Slit
5
15
25
40
3 days 7 days
5 15 25 40
0.0
0.1
0.2
5.0
10.0
5 15 25 40
0.0
0.5
1.0
5 15 25 40
0
2
4
6
8
10
5 15 25 40
0.0
0.1
1.0
2.0
3.0
4.0
12
• Manure application method & timing‐relative to rain events?
distance from the slit (cm)
Subsurface injection
Surface
application
Surface
application
Surface
application
distance from the slit (cm)
Subsurface injection
distance from the slit (cm)
Subsurface injection
CECs in soil
Days between manure application & rain event – after rain
Concentration(ng/g,d.w.)

13. Losswithrunoffwater/initialamountinmanure(%)
0
1
2
3
Surface application
Subsurface injection
Losswithrunoffwater/initialamountinmanure(%)
0.00
0.10
0.20
0.30
0.40
0.50
Pirlimycin
0 3 7
0.00
0.05
0.10
0.15
Sulfamerazine
Chlortetracycline
Days between manure application and rain event
0 3 7
0.0
0.2
0.4
0.6
0.8
Tylosin
13
• Manure application method & timing relative to rain events?
Loss of CECs with surface runoff

14. Losswithrunoffwater/initialamountinmanure(%)
0
1
2
3
Surface application
Subsurface injection
Losswithrunoffwater/initialamountinmanure(%)
0.00
0.10
0.20
0.30
0.40
0.50
Pirlimycin
0 3 7
0.00
0.05
0.10
0.15
Sulfamerazine
Chlortetracycline
Days between manure application and rain event
0 3 7
0.0
0.2
0.4
0.6
0.8
Tylosin
14
• Manure application method & timing relative to rain events?
Loss of CECs with surface runoff

15. Losswithrunoffwater/initialamountinmanure(%)
0
1
2
3
Surface application
Subsurface injection
Losswithrunoffwater/initialamountinmanure(%)
0.00
0.10
0.20
0.30
0.40
0.50
Pirlimycin
0 3 7
0.00
0.05
0.10
0.15
Sulfamerazine
Chlortetracycline
Days between manure application and rain event
0 3 7
0.0
0.2
0.4
0.6
0.8
Tylosin
15
• Manure application method & timing relative to rain events?
Loss of CECs with surface runoff

16. 16
• Spatial variation?
Concentration(ng/g,dryweightbasis)
0
50
100
150
200
250
300
350
400
Pirlimycin
Sulfamerazine
Tylosin
Chlortetracycline
Days between manure application rain - after rain
0 day
0-5 cm
Surface
Slit
5
15
25
40
0-5 cm
5-20 cm
5 15 25 40
0.0
0.1
0.2
5.0
10.0
5 15 25 40
0.0
0.5
1.0
Surface
Slit
5
15
25
40
0
50
100
150
200
250
300
350
5-20 cm
Surface
Slit
5
15
25
40
3 days 7 days
5 15 25 40
0.0
0.1
0.2
5.0
10.0
5 15 25 40
0.0
0.5
1.0
5 15 25 40
0
2
4
6
8
10
5 15 25 40
0.0
0.1
1.0
2.0
3.0
4.0
distance from the slit (cm)
Subsurface injection
Surface
application
Surface
application
Surface
application
distance from the slit (cm)
Subsurface injection
distance from the slit (cm)
Subsurface injection
CECs in soil
Concentration(ng/g,d.w.)

17. 17
• Temporal change?
Time since rain event (days)
rain 5 14 60 180
Concentrationofchlortetracycline(ng/g,d.w.)
0
50
100
150
200
250
Days between manure application & rain event:
injection slit
Time since rain event (days)
rain 5 14 60 180
Concentrationofchlortetracycline(ng/g,d.w.)
0
10
20
30
40
50
Surface application
0 3 7
Chlortetracycline in soil decreased with time since rain event

18. • Manure subsurface injection concentrates CECs within the
injection slits
 Conclusions
• Loss of CECs in surface runoff occurred during the 1st rain
event after manure application
• Apply manure at lease 3 days before a major rain event
• Minor horizontal transfer of CECs from the injection slits
• Less CECs left the field with surface runoff when manure is
subsurface injected comparing to surface application
18

19.  Looking forward
Regional scale observation?
19
Chesapeake bay watershed
Region (no‐till + reduced tillage)/Crop land
United States 36%
Mid‐South &
Great Plains
30‐55%
Chesapeake Bay
Watershed
> 80%

20.  Looking forward
Antimicrobial resistant development?
20

21. Acknowledgement
• USDA‐NIFA award #2013‐67019‐21355
• Virginia Tech College of Agriculture and Life Sciences
21

22. Thank you!
Questions?
22