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Addition of Condensed TanninSources to Broiler Litter BeforeDeep-StackingZ. S. Wang a , A. L. Goetsch b , K. K. Park a , A. R.Patil a , B. Kouakou a , D. L. Galloway Sr. a & J. E.Rossi aa Department of Animal Science, University ofArkansas, Fayetteville, AR, 72701, USAb South Central Family Farm Research Centre,USDA-ARS, 6883 So. St. Hwy 23, Booneville, AR,72927-9214, USAPublished online: 11 Nov 2011.

To cite this article: Z. S. Wang , A. L. Goetsch , K. K. Park , A. R. Patil , B. Kouakou ,D. L. Galloway Sr. & J. E. Rossi (1996) Addition of Condensed Tannin Sources to BroilerLitter Before Deep-Stacking, Journal of Applied Animal Research, 10:1, 59-79, DOI:10.1080/09712119.1996.9706131

To link to this article: http://dx.doi.org/10.1080/09712119.1996.9706131

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J. Appl. Anim. Res. 10 (1996) : 59-79

Addition of Condensed Tannin Sources to Broiler Litter Before Deep-Stacking132,3

Z.S. Wang, A.L. Goetsch4, K.K. Park, A.R. Patil, B. Kouakou, D.L. Galloway, Sr., J.E. Rossi

Department of Animal Science, University of Arkansas, Fayetteville,

AR 72701, USA and

South Central Family Farm Research Centre USDA-ARS, 6883 So. St. Hwy 23, Booneville,

4

AR 72927-9214, USA

(Received February 22, 1996; accepted May 25, 1996)

Abstract Wang, Z.S., Goetsch, A.L., Park, KK, Patil, A.R., Kouakou, B., Galloway, D.L., Sr. and Rossi, J.E. 1996. Addition of condensed tannin sources to broiler litter before deep-stacking. J. Appl. Anim. Res., 10: 59-79.

Different sources and levels of condensed tannins were thoroughly mixed (M) or covered (C) with broiler litter in plastic vessels fitted with a dacron top. Containers were placed in a deep-stack for 3 or

'Published with the approval of the' Director of the Arkansas Agricultural Experiment Station, Manuscript Number 96009. 'Appreciation is expressed to the NRI Competitive Grants ProgramRTSDA (Award Number 93-37500-9152) for partial financial support, and to Jim Hornsey (Planters-Lifesavers, Fort Smith, AR), Northrup King (New Deal, TX), Tannin Corp. (Peabody, MA) and Larry Morrison (Monett, MO) for providing materials. 3Mention of a trademark or proprietary product in this paper does not constitute a guarantee or warranty of the product by the USDA or the ARS and does not imply its approval to the exclusion of other products that may be suitable. 4 F ~ r reprint requests-Phone : 501/675-3834; Fax : 501/675-2940.

59

J. Appl. Anim. Res. 0971-2119/96/$05.00 8 GSP, India

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60 Z.S. Wang and coworkers

9 wk to determine effects on constituent concentrations, recoveries and in aitu ruminal disappearances. Added substrates were commercial vegetable extract WE; quebrach), peanut skins (PS), bird-resistant sorghum grain (BS) and regular sorghum grain (S), with condensed tannin (catechin equivalents) concentrations of 68.4, 24.7, 0.6 and O.O%, respectively. Levels (total dry matter basis) were 0, 0.04, 0.09, 0.17, 0.35, 1.3, 2.6, 5.1 and 10.3% VE; 0, 3.6, 7.1, 14.2 and 28.4% PS; and 0, 5, 10, 20 and 40% BS and S. Condensed tannin recovery for VE [DO, 113, 105, 98, 63, 27, 18, 12 and 10% for M and 125, 118, 107, 84, 59, 37, 23, 20 and 23% for C at 3 wk (SE 2.8); 87, 74, 66, 61, 44, 22, 16, 9 and 11% at 9 wk (SE 3.3) with 0, 0.04, 0.09, 0.17, 0.35, 1.3, 2.6, 5.1 and 10.3% VE, respectively], PS [139, 29, 18, 14 and 20% for M and 126, 37, 27, 25 and 46% for C at 3 wk (SE 4.9); 52, 21, 12, 9 and 6% for M and 91, 23, 14, 10 and 17% for C at 9 wk (SE 4.9) with 0, 3.5, 7.1, 14.2 and 28.4% PS,respectively] and BS 1135, 99, 109, 75 and 50% at 3 wk and 97, 88, 88, 68 and 48% at 9 wk (SE 5.1) with 0, 5, 10, 20 and 40% BS, respectively] generally was less for 9 us 3 wk of deep-stacking and decreased with increasing level of condensed tannin source. Addition of condensed tannin sources did not markedly alter concentrations of nitrogen fractions, recoveries of organic matter or nitrogen or in situ dry matter or nitrogen disappearances. In conclusion, addition of condensed tannin sources to broiler litter before deep-stacking decreased recovery of assayable condensed tannins, with effects varying with level and method of addition.

Key words : Broiler litter, condensed tannins, deep-stack, peanut skins, sorghum grain.

Introduction Tannins are high molecular weight polyphenolic compounds of plant origin classified as hydrolyzable (polymers of gallic acid and various sugars) or condensed (polymers of flavenoid phenols; Kumar and Singh, 1984). A variety of feedstuffs contain condensed tannins, the nature of which varies considerably among feedstuffs (Kumar and Singh, 1984; Robbins et al., 1987; Jansman, 1993; Reed, 1995). Examples of feedstuffs high in condensed tannins include some vegetables (Jansman, 1993), peanut skins (Hale and McCormick, 1981; McBrayer et al., 1983) and bird-resistant sorghum grain (Mitaru et al., 1984). Typically, only limited amounts of feedstuffs high in condensed tannins are consumed by ruminants because

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Condensed tannins and broiler litter 61

of protein binding and precipitation, which interefere with digestion and (or) impair palatability (Kumar and Vaithiyanathan, 1990; Jansman, 1993; Reed, 1995). Therefore, inexpensive means to avert or lessen adverse effects of condensed tannins on ruminant digestion and feed intake are of interest.

Broiler litter is abundant and relatively inexpensive in many parts of the world. Frequently broiler litter is processed for feeding to ruminants by a method known as deep-stacking. Conditions in broiler litter deep-stacks include 20 to 30% moisture, a pH of 7 to 9 and temperature of 40 to 60C (Ruffin and McCaskey, 1990). Ammonia in broiler litter is approximately 14% of total nitrogen; uric acid initially contributes a greater proportion of total nitrogen (e.g., 30%; Fontenot and Jurubescu, 1980), although its concentration declines with conversion to urea and ammonia (Oltjen and Dinius, 1976).

Various treatments of high-tannin sorghum grain decrease assayable condensed tannin concentration, increasing nutritional value and lessening or eliminating feeding limits, although exact modes of action are unclear (Reichert et al., 1980). Ammonia reacts with condensed tannins, involving binding or chemical changes of functional groups necessary for protein binding (Price et al., 1978a). However, Price et al. (1979) concluded that moist alkaline conditions, rather than an effect of ammonia per se, were responsible for decreases in assayable condensed tannins and increased feeding value. Effectiveness of such treatments appears enhanced by increased temperature (e.g., 60 us 25C; Russell and Lolley, 1989) and moisture concentration (e.g., 25 > 15%; Mitaru et al., 1984). Hence, exposing feedstuffs containing condensed tannins to conditions in a broiler litter deep-stack might decrease condensed tannin concentration and improve nutritive value. In this regard, Patil et al. (1993) increased nitrogen and decreased condensed tannin concentrations by deep-stacking a mixture of peanut skins and broiler litter compared with broiler litter alone, suggesting that changes resulted from ammonia binding to and cleavage of condensed tannins (Kumar and Singh, 1984; Hill et al., 1986). Therefore, objectives of this experiment were to determine effects on nutrient composition and recovery of addition of different sources and various levels of condensed. tannins by thorough mixing or covering with broiler litter before deep-stacking for 3 or 9 wk.

Materials and Methods Broiler litter was obtained from a commercial production unit after six growing periods, each approximately 42 d in length. Initial moisture

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62 Z.S. Wang and coworkers

concentration was 20.8%. Bedding was a mixture of pine shavings and rice hulls applied before the first growing period; packed litter was broken up and mixed with other litter after each growing period. An aliquot of litter passing through a 4.3 x 1.9 cm screen was used to construct substrates for incubation.

Substrates (250 g dry matter and 25% moisture) were constructed and placed in duplicate l-L plastic beakers (11.1 cm top id., 8.9 cm bottom id . and 14.3 cm tall) with a dacron cloth top (50 to 75 micron pores). Vessels were placed in the center of a 1.4 x 2.0 m deep-stack, approximately 1.6 m tall and at 25% moisture, of the same broiler litter used for substrate preparation. Deep-stacking was for 3 or 9 wk. Perforated polyvinylchloride pipes (7.0 cm i.d. and 7.6 cm o.d.), open at one end, were placed at the bottom of the bay to promote oxygen entry.

Substrates consisted of different levels of broiler litter and commercial vegetable extract [ 0%) cold soluble quebracho spray-dried powder; Tannin Corp. , Peabody, MA], peanut skins (PS; Planters- Lifesavers, Fort Simth, AR), ground (2-mm screen) bird-resistant sorghum grain [(BS) Savanna 5 ; Northrup King, New Deal, TX] or ground (2-mm screen) regular sorghum grain [(S) KS 7144; Northrup King, New Deal, TX]. Substrates were comprised of 0, 0.04, 0.09, 0.17, 0.35, 1.3, 2.6, 5.1 and 10.3% VE; 0, 3.6, 7.1, 14.2 and 28.4% PS; and 0, 5 , 10, 20 and 40% BS and S. Levels of added condensed tannins for 3.6, 7.1, 14.2 and 28.4% PS were similar to those for 1.3, 2.6, 5.1 and 10.3% VE respectively. Likewise, condensed tannin additions with 5, 10, 20 and 40% BS were comparable to those for 0.04, 0.09, 0.17 and 0.35% VE, respectively. Substrate ingredients were thoroughly hand-mixed in a plastic bag before placement in beakers, or broiler litter was placed at the bottom of the beaker and covered with VE, PS, BS or S.

Temperature was measured daily for 21 d and once weekly thereafter with a commercial thermometer placed in the center of the deep-stack. After removal of vessels, substrates were hand-mixed. Substrate pH was measured subsequent to mixing 20 g with 50 mL of deionized water. Remaining substrate was stored at -132, later thawed and ground to pass a 2-mm screen.

Substrates were analyzed for dry matter, ash, Kjeldahl nitrogen (N; Association of Official Analytical Chemists, 1984), acid detergent fiber N (Goering and Van Soest, 1970) and soluble N (0.15 M NaC1; Waldo and Goering, 1979). Ammonia N concentration was determined @roderick and Kang, 1980) after placing 1 g of substrate in 100 mL of 0.15 M NaCl and

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Condensed tannins and broiler litter 63

filtering, and soluble protein was measured by the procedure of Prigge et al. (1976). Nonprotein N and nonammonia nonprotein N concentrations were determined by difference. The concentration of catechin equivalents was determined by the moditied vanillin-HC1 method of Price et al. (197813).

In situ ruminal dry matter and N disappearances were determined for 0, 0.17, 2.6 and 10.3% VE, 0, 7.1 and 28.4% PS, and 0 and 20% BS and S. A ruminally cannulated beef steer consuming long-stemmed bermudagrass hay at 1.6% of body weight (dry matter basis), with water and trace mineralized salt available free-choice was used for incubation. Duplicate dacron bags (53ilO micron pores; 8 x 5 cm) containing 1 g of substrate were incubated for 12 or 48 h. Upon removal, bags were washed by hand until rinse fluid was clear and residue was analyzed for dry matter and N. Additional bags incubated for 48 h were subjected to pepsin digestion (Association of Official Analytical Chemists, 1984) before N analysis. Predicted intestinal digestion of N was estimated as the difference between residual N after 12 h of ruminal incubation and residual pepsin insoluble N at 48 h. In addition to treatment substrates, broiler litter, VE, PS, BS and S sampled before deep-stacking were subjected to aforementioned procedures to estimate recovery or net change in concentration with deep-stacking (i.e., difference between final or actual and initial or expected concentration).

Data were analyzed separately for VE, PS, BS and S substrates by the General Linear Models procedure of Statistical Analysis System (1990), first with a full model containing level of non-broiler litter substrate ingredient, length of deep-stacking, method of addition and all interactions thereof. The analysis was conducted separately for each length of deep-stacking when the three-way interaction was significant (P<0.05) and the three-way interacation was removed from the model when nonsignificant (P>0.05). Differences among means were determined by least significant difference procedures when overall F values were significant (P<0.05). Data were presented in tabular form in the simplest manner deemed possible. That is, with nonsignificant interactions main effects were listed. Concommitantly, means for the various treatment combinations appear when interactions occurred; means for variables with nonsignificant effects are not listed in tables; and mode of presentation was not consistent among variables. Because of the high number of means presented in tables, superscripts denoting significant differences between individual treatment means were not listed.

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Tab

le 1

C

ompo

sitio

n of

sub

stra

te in

gred

ient

s Q

, Ip

Item

B

roile

r V

eget

able

Pe

anut

B

ird-

resi

stan

t R

egul

ar

litt

er

extr

act

skin

s so

rghu

m g

rain

so

rghu

m g

rain

Org

anic

mat

ter

(% o

f DM)

Tot

al n

itrog

en (

% o

f DM)

Solu

ble

N (

% o

f to

tal

N)

75.2

86

.8

97.3

98

.2

98.3

4.98

0.

19

2.43

2.

11

1.86

61.4

10

0.0

7.8

12.3

17

.3

Aci

d de

terg

ent f

iber

N (9% o

f to

tal

N)

7.9

20.3

18

.6

20.6

19

.5

Inso

lubl

e av

aila

ble

N (

% o

f to

tal

N)

30.7

0.

0 73

.6

67.1

63

.2

Solu

ble

true

pro

tein

N (

% o

f to

tal

N)

12.8

65

.5

5.8

8.9

12.7

N

onpr

otei

n N

(%

of

tota

l N

) 48

.6

34.5

2.

0 3.

4 4.

6

Am

mon

ia N

(%

of

tota

l N

) 6.

7 0.

0 0.

0 0.

0 0.

0 N

onam

mon

ia n

onpr

otei

n N

('3%

of t

otal

N)

41.9

34

.5

2.0

3.4

4.G

Con

dens

ed t

anni

ns2

(% o

f DM)

0.2

68.4

24

.7

0.6

0.0

In situ DM d

isap

pear

ance

, 12

h (%

) 70

.0

48.8

68

.3

65.0

In situ DM d

isap

pear

ance

, 48

h (

%)

In situ

N d

isap

pear

ance

, 12

h (

%)

85.0

88.1

91.9

In situ

N d

isap

pear

ance

, 48

h (

%)

Peps

in i

nsol

uble

N a

fter

48

h of

77.5

94

.2

95.0

36.4

41

.8

43.7

54

.6

75.3

75

.9

rum

inal

incu

batio

n (%

of

tota

l N

) 4.

4 35

.9

4.4

2.4

Pred

icte

d in

test

inal

N d

iges

tion3

(%

of

tota

l N

) 3.

7 9.

5 20

.3

21.7

'DM

= dr

y m

atte

r; N=

nitr

ogen

. 'C

atec

hin

equi

vale

nts.

3Diff

eren

ce b

etw

een

resi

dual

N a

fter

12 h

of

rum

inal

incu

batio

n an

d re

sidu

al p

epsi

n in

solu

ble

N af

ter 4

8 h

of r

umin

al in

cuba

tion.

N

YJ,

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Condensed tannins and broiler litter 65

Results and Discussion Temperature and pH Temperature in the center of the deep-stack was 51, 57, 57, 50, 50, 46, 44, 42 and 41C in wk 1, 2, 3, 4, 5, 6, 7, 8 and 9, respectively. The pH for VE and S substrates was not affected by treatments but was greater (P<0.05) at 9 than 3 wk of deep-stacking for PS (7.66 us 7.25; SE 0.093) and BS substrates (7.64 us 7.32; SE 0.106).

Substrate ingredient composition Composition of substrate ingredients is given in Table 1. Ammonia N concentration in broiler litter was slightly lower than an average of 14% in litter noted by Fontenot and Jurubescu (1980). The VE condensed tannin concentration was 68%, which is only slightly less than the condensed tannin concentration of 72 to 75% indicated by the manufacturer. The concentration of condensed tannins in PS was greater than observed by others (16 to 20% : Hale and McCormick, 1981 and McBrayer et al., 1983; 13% : Patil et al., 1993). Condensed tannin concentration in BS was considerably less than expected (Jansman, 1993), although condensed tannins were not detected in S. In situ dry matter and N disappearances for broiler litter were similar to results of Mandebvu et al. (1995a,b), Park et al. (1995) and Wang et al. (1996). In situ disappearances of the two sorghum grains were similar, possibly reflecting the moderate or low level of condensed tannins in BS. Vegetable extract was not subjected to in situ incubation because of its soluble nature.

Vegetable extract Organic matter recovery for mixed substrates did not markedly vary with VE level, although that for covered substrates decreased slightly with increasing level of VE (Table 2). Perhaps covering broiler litter with VE affected microbial activity (Reed, 1995) in litter beneath relatively more than when thoroughly mixed with litter. As in our experiment, organic matter recovery slightly greater than 100% was noted in previous experiments with similar methodology, which may relate to influx of microorganisms into substrate vessels from surrounding broiler litter in the deep-stack (Mandebvu et al., 1995a,b; Park et ~1.~1995; Wang et al., 1996). Organic matter recovery was slightly less (P<0.05) for deep- stacking 9 us 3 wk (101.8 us 102.4%; SE 0.10).

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Q)

Tab

le 2

Q

)

Effects o

f le

vel o

f ve

geta

ble

extr

act

(que

brac

ho)

and

met

hod

of a

dditi

on to

bro

iler

litt

er o

n co

nstit

uent

rec

over

y an

d pe

rcen

tage

unit

chan

ge i

n co

ncen

trat

ion

of c

onst

ituen

ts a

nd in s

itu d

isap

pear

ance

aft

er d

eep-

stac

king

for

3 or

9 wk

~

Veg

etab

le e

xtra

ct l

evel

(% o

f to

tal

dry

mat

ter)

It

em'

SE

0 0.

04

0.09

0.

17

0.35

1.

3 2.

6 5.

1 10

.3

Org

anic

mat

ter

reco

very

(%)

Mix

ed2

101.

9 10

3.2

102.

2 10

3.0

102.

5 10

2.8

102.

5 10

2.5

102.

5 co

vere

d2

103.

0 10

3.1

102.

9 10

2.1

101.

3 10

1.2

100.

1 10

0.0

100.

6 0.

30

Y Total

N r

ecov

ery

(%)

Mix

ed

95.0

94

.2

93.7

94

.2

92.4

95

.9

94.9

95

.9

97.3

!n

Cov

ered

91

.9

94.6

93

.1

91.2

97

.1

92.7

95

.9

94.8

98

.1

1.11

3

Solu

ble

N c

hang

e (%

') G i

Aci

d d

eter

gen

t fi

ber

N c

hang

e (%

') s i+

Mix

ed

-4.4

-2

.4

-5.4

-3

.2

-8.7

-4

.7

-6.3

-6

.8

-8.5

C

over

ed

-5.8

-4

.4

-7.0

-3

.5

-2.3

-3

.7

-4.0

-6

.3

-4.3

0.

96

8

Mix

ed

1.5

1.4

1.0

1.1

1.3

1.4

2.6

2.6

4.1

Cov

ered

1.

3 1.

7 1.

1 1.

4 0.

4 1.

4 1.

2 1.

6 1.

8 0.

37

3-w

k le

ngth

1.

1 1.

1 0.

8 1.

4 1.

1 1.

3 1.

1 1.

3 1.

5 9-

wk

leng

th

1.6

1.9

1.3

1.1

0.6

1.5

2.8

2.9

4.3

0.37

In

solu

ble

avai

lab

le N

cha

nge

(%')

Mix

ed

2.9

0.9

4.4

2.0

7.4

3.3

3.7

4.2

4.4

Cov

ered

4.

5 2.

7 5.

0 2.

1 1.

9 2.

3 2.

8 4.

7 2.

5 0.

85

a

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Solu

ble

tru

e p

rote

in N

cha

nge

(%'I

Mix

ed

-3.9

C

over

ed

-8.1

N

onpr

otei

n N c

hang

e (%

') 3-

wk

leng

th

Mix

ed

2.3

Cov

ered

-2

.4

9-w

k le

ngth

1.

8 A

mm

onia

N c

hang

e (%

'09)

-1.6

N

onam

mon

ia n

onpr

otei

n N

cha

nge

(%3)

9-w

k le

ngth

1.

4 C

hang

e in

in s

itu

12-

h N

pi

sapp

eara

nce (%3~

0.3

3-w

k le

ngth

3.

5

-0.3

-0

.2

1.7

-3.7

-0

.8

-2.4

-6

.7

-4.5

-0

.6

-3.2

-4.0

-4

.6

-2.4

-0

.1

-1.4

-2

.5

-4.6

-3

.5

-3.4

-4

.4

-0.9

1.

2 1.

0 -5

.0

-1.6

-1

.3

0.6

1.0

2.3

2.7

0.2

-2.8

-2

.3

-2.3

-2

.0

-1.6

-2

.9

-3.6

-9

.0

-7.9

-0.7

C

hang

e in

pep

sin

inso

lubl

e N

aft

er 4

8-h

rum

inal i

ncub

atio

n, 9

-wk

leng

th (%

') M

ixed

1.

3 1.

3 C

over

ed

1.9

2.3

Cha

nge

in p

redi

cted

inte

stin

al N

dig

esti

on, 9

-wk

len

gth

(%')

Mix

ed

0.3

-0.7

C

over

ed

-0.9

-1

.1

Con

dens

ed ta

nnin

' re

cove

ry (%

) 3-

wk

leng

th

Mix

ed

119.

6 11

2.8

104.

7 98

.4

63.3

27

.2

Cov

ered

12

4.7

118.

1 10

7.1

84.0

59

.3

37.4

0.3

-2.8

-6.1

-3

.7

-2.8

2.

9

-5.5

-8

.0

-1.1

2.8

1.9

-0.7

-0

.5

18.4

23

.2

-5.1

-0

.6

-1.8

-2

.5

-5.3

1.

9

-1.6

-9

.6

12.4

19

.7

0.2

-3.1

-7.2

-3

.2

-4.6

2.

7

-5.9

-9

.4

-1.7

5.2

1.9

-3.0

-1

.1

10.2

22

.5

1.66

1.17

1.

31

0.51

1.25

0.43

0.44

0.39

2.79

9-

wk

leng

th

86.7

73

.5

66.2

61

.3

43.7

22

.3

15.9

9.

3 11

.4

3.29

*N =

nitr

ogen

. 'V

eget

able

ext

ract

and

bro

iler

litt

er w

ere

thor

ough

ly m

ixed

, or

vege

tabl

e ex

trac

t w

as p

lace

d ab

ove

broi

ler

litt

er in

sep

arat

e la

yers

. 'P

erce

ntag

e un

it c

hang

e (f

inal

min

us i

nitia

l).

'Cat

echi

n eq

uiva

lent

s.

Dow

nloa

ded

by [

Mos

kow

Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

60 Z.S. Wang and ooworkers

No consistent effects of VE level or method of addition on N recovery were observed (Table 2). Solubility of N was slightly less after than before deep-stacking, although consistent effects of VE level or differences between addition methods did not occur. Magnitude of change in soluble N concentration was greater (P<0.05) after 9 than 3 wk of deep-stacking (-5.8 us -4.4%; SE 0.32). Two-way interactions in change in acid detergent fiber N concentration were observed between VE level and incubation length and method of addition. It appeared that acid detergent fiber N concentration in mixed substrates and after 9 wk of deep-stacking was increased slightly by increasing the level to 12.6% of total DM. However, corresponding changes in concentration of insoluble available N were not observed. Change in soluble true protein was negative, although differences between methods of addition or among levels of VE were inconsistent. Likewise, implications of significant interactions and treatment effects in change in nonprotein N concentration were not apparent.

Ammonia N concentration was slightly less than expected with 0 and 0.04% VE but greater for VE levels 10.35% (Table 2). Change in nonammonia nonprotein N concentration was positive for 0% VE at 3 and 9 wk but negative for ~ 0 . 0 9 % VE. Magnitude of change for these VE levels was generally greater at 9 us 3 wk of deep-stacking. Changes in concentrations of ammonia and nonammonia nonprotein N may reflect that, particularly at 9 wk, VE addition increased ammonia formation from nonammonia nonprotein nitrogenous compounds (i.e., primarily uric acid and urea) without markedly altering ammonia concentration. Thus, condensed tannins in VE may not have facilitated retention of all ammonia initially present or formed in litter. Level of VE, addition method and length of deep-stacking had no or relatively small effects on change in in situ dry matter and N disappearances and residual pepsin insoluble N following in situ ruminal incubation. Magnitude of change in in situ N disappearance with 48 h of ruminal incubation was greater (P<0.05) at 9 us 3 wk of deep-stacking (1.4 us 0.8%; SE 0.13).

Recovery of condensed tannins at 3 wk of deep-stacking was greater than 100% for 0, 0.04 and 0.09% VE, although recovery generally decreased as VE level increased form 0 to 10.3% of total dry matter (Table 2). High recovery of condensed tannins in substrates with low initial concentrations may have resulted from analytical problems involving reaction of non-tannin substances with vanillin (Burns, 1971). This procedure can overestimate concentration of condensed tannins in samples with low concentrations relatively more than do other

Dow

nloa

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by [

Mos

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Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

Condensed tannins and broiler litter 69

procedures (Maxson and Rooney, 1972). For substrates deep-stacked for 3 wk, condensed tannin recovery at all but two VE levels was less for mixed than for covered substrates, possibly indicative of greater exposure of condensed tannins in mixed substrates to ammonia or moist alkaline conditions. Condensed tannin recovery was less at 9 us 3 wk of deep-stacking and at 9 wk of deep-stacking recovery was similar between addition methods. This suggests that prolonged exposure to deep-stack conditions of condensed tannins in covered substrates compensated for possibly less exposure per unit time when mixed. In accordance, Price et al., (1979) observed that dilute alkali treatments of BS required longer treatment times than with more concentrated solutions. It appears that rate of condensed tannin deactivation in our experiment may have been lower than rates for reconstituted BS with added urea of 44 and 89% per day at 25 and 60 C, respectively, noted by Russell and Lolley (1989).

Peanut skins Organic matter recovery was not influenced by level of PS and was greater (P<0.05) for mixed than for covered substrates (Table 3). An interaction (P<0.05) in N recovery between deep-stacking length and PS level was observed, which appeared primarily the result of relatively high recovery for 28.4% PS with 9 wk of deep-stacking. Patil et al. (1993) observed an effect of mixing broiler litter with PS (27% of total dry matter) in a deep-stack. Reasons for comparatively little effect of PS addition on N recovery in our experiment are unclear but could involve use of small quantities of substrate contained in plastic beakers with a dacron cloth top rather than employment of an entire deep-stack. The difference between lengths of deep-stacking in change in soluble N concentration was opposite that noted for VE substrates. Likewise, magnitude of change in soluble N concentration was greater (P<0.05) for mixed us covered substrates; whereas, differences between addition methods were inconsistent among substrate VE levels.

As noted for VE substrates, change in acid detergent fiber N concentration between that before and after deep-stacking was greater for high PS levels compared with lower levels (Table 3). Change in acid detergent fiber N concentration with high PS levels was greater (P<0.05) for mixed us covered substrates and for all PS levels a t 9 us 3 wk deep-stacking. Factors responsible for these interactions are unclear. For both VE and PS substrates, generally corresponding changes in residual pepsin insoluble N were noted. However, change in predicted intestinal N digestion was not appreciably altered by VE or PS levels. Perhaps binding of condensed tannins to proteins, quite possibly of microbial origin, were

Dow

nloa

ded

by [

Mos

kow

Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

4

0

Tab

le 3

E

ffec

ts o

f le

vel o

f pe

anut

ski

ns a

nd m

etho

d of

add

ition

to

broi

ler

litt

er o

n co

nstit

uent

rec

over

y an

d pe

rcen

tage

un

it c

hang

e in

con

cent

ratio

n of

con

stitu

ents

and

in

situ

dis

appe

aran

ce a

fter

dee

p-st

acki

ng f

or 3

or

9 wk

Add

ition

met

hod'

Pe

anut

ski

ns l

evel

(% o

f to

tal dr

y m

atte

r)

0 3.

6 7.

1 14

.2

28.4

SE

3

wk

9 w

k SE

Mixed

Cov

ered

SE

In

cuba

tion

leng

th

Item

Org

anic

mat

ter

reco

very

(%

)

Total

N r

ecov

ery

(%)

3-w

k le

ngth

94

.1

93.5

9-

wk

leng

th

93.6

91

.8

Solu

ble

N c

han

ge (%

') A

cid

det

erge

nt

fib

er N

ch

ange

(%')

Mixed

0.9

1.7

Cov

ered

0.

9 1.

8 3-

wk

leng

th

0.7

0.8

9-w

k le

ngth

1.

2 2.

7 In

solu

ble

avai

lab

le N

cha

nge

(%')

3-w

k le

ngth

3.

2 3.

0

Solu

ble

true

pro

tein

N

chan

ge (%

') N

onpr

otei

n N

ch

ange

,

Ammo

nia

N c

hang

e (%

'I 1.

7 3.

0

9-w

k le

ngth

3.

9 -0

.3

9-wk le

ngt

h (%

'I -6

.0

-4.2

102.

4 10

0.5

0.18

94.0

97

.2

98.4

96

.8

95.6

10

3.2

1.03

-3

.8

-2.4

0.

49

-4.0

-2

.2

0.49

p

!n

2.5

4.7

5.6

3 2.

0 2.

9 3.

0 0.

34

Gz E

2.0

2.5

3.3

2.5

5.1

6.2

0.34

3.0

0.9

-0.6

-0

.7

-2.4

-6

.3

0.96

-2.5

0.

1 0.

75

-2.9

0.

5 0.

75

-3.3

0.

7 0.

3 1.

14

-1.4

-3.6

0.72

3.

1 3.

4 5.

0 0.

52

2.2

4.2

0.33

4 F 2

Dow

nloa

ded

by [

Mos

kow

Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

Con

dens

ed t

anni

n'

reco

very

(%

) 3-

wk

leng

th

Mix

ed

139.

1 29

.2

18.2

14

.0

19.6

C

over

ed

126.

3 37

.0

26.9

25

.2

45.8

4.

95

9-w

k le

ngth

M

ixed

52

.0

20.8

11

.5

8.9

6.0

8

Cov

ered

91

.1

23.4

14

.4

10.2

16

.8

4.91

E

Cha

nge

in i

n s

itu

12-

h N

8. F :. E. -3

.5

-1.8

0.

34

disa

ppea

ranc

e (%

') -0

.4

-1.9

-5

.7

0.41

disa

ppea

ranc

e (%

') -0

.4

-2.1

-5

.4

0.40

-3

.3

-2.0

0.

33

Cha

nge

in i

n S

ifU

48-h N

Cha

nge

in p

epsi

n i

nso

lub

le N

aft

er 4

8-h

ru

min

al in

cuba

tion

(%3)

3-w

k le

ngth

a

Mix

ed

0.2

2.5

7.3

2 C

over

ed

0.4

1.9

4.7

0.13

9-

wk

leng

th

1.5

3.2

7.5

0.47

IN

-nit

roge

n 'P

eanu

t sk

ins

and

broi

ler

litte

r w

ere

thor

ough

ly m

ixed

, or

pean

ut s

kins

wer

e placed a

bove

bro

iler

litte

r in

sep

arat

e la

yers

. 3P

e~en

tage

unit

cha

nge

(fin

al m

inus

ini

tial

). 4C

atec

hin

equi

vale

nts.

W

c = - K

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nloa

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by [

Mos

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Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

4

Tab

le 4

N

Eff

ects

of

leve

l of

bird

-res

ista

nt s

orgh

um g

rain

and

met

hod

of a

dditi

on t

o br

oile

r li

tter

on

cons

titue

nt r

ecov

ery

and

perc

enta

ge u

nit

chan

ge i

n co

ncen

trat

ion

of c

onst

ituen

ts a

nd in

situ

dis

appe

aran

ce a

fter

dee

p-st

acki

ng f

or

3 or

9 wk

Item

' B

ird-

resi

stan

t so

rghu

m g

rain

In

cuba

tion

leng

th

Add

ition

met

hod2

(%

of

tota

l dr

y m

atte

r)

0 5

10

20

40

SE

3 w

k 9

wk

SE

Mix

ed C

over

ed

SE

Org

anic

mat

ter

reco

very

(%)

101.

7 10

1.6

101.

6 10

1.2

100.

5 0.

23

101.

6 10

1.0

0.14

10

2.1

100.

5 0.

14

Tot

al N

rec

over

y (%

) 95

.5

93.5

0.

58

3-w

k le

ngth

93

.8

93.2

96

.6

91.6

95

.3

ol

9-w

k le

ngth

90

.9

92.9

95

.7

95.8

99

.4

1.29

3

Solu

ble

N c

hang

e (%

') -2

.1

-3.7

0.

39

z 8 A

cid

det

erge

nt

fib

er N

Inso

lubl

e av

aila

ble

N

Non

prot

ein

N c

han

ge (%

')

chan

ge (%

'I 0.

3 1.

7 2.

0 3.

3 4.

2 0.

40

1.9

2.7

0.25

chan

ge (%

') 3.

7 1.

5 -0

.4

-0.5

-1

.7

0.71

9 2

3-w

k le

ngth

Covered

1.6

0.3

3.0

4.4

4.6

1.44

9-

wk

leng

th

-1.9

1.

0 0.

25

Mixed

-0.7

-3

.4

-0.2

12

.4

6.9

Am

mon

ia N

ch

ange

(%3)

2.1

2.4

1.9

2.4

3.9

0.42

1.

6 3.

5 0.

27

Dow

nloa

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by [

Mos

kow

Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

Non

amm

onia

non

pro

tein

N c

han

ge (%

') 3-

wk

leng

th

Mix

ed

-2.5

-3

.9

-1.2

8.

3 4.

4 C

over

ed

1.3

-1.6

1.

1 3.

0 2.

2 1.

16

9-w

k le

ngth

Mixed

-7.8

-4

.9

-5.5

-4

.5

-3.8

C

over

ed

-0.4

-6

.2

-2.3

-1

.8

-1.9

1.

18

Cha

nge

in in

sit

u 1

2-h

N

Cha

nge

in in

eit

u 4

8.h

N

Cha

nge

in p

epsi

n in

solu

ble

N a

fter

48-

h r

umin

al in

cuba

tion,

9-wk le

ngt

h (%

')

dis

app

eara

nce

(%3)

-0

.3

-2.3

0.

34

dis

app

eara

nce

(%')

-0.7

-3

.8

0.57

3-w

k le

ngth

0.

9 2.

3 9-

wk

leng

th

1.6

1.5

0.25

9-w

k le

ngth

(%')

-0.9

2.

4 0.

36

Cha

nge

in p

red

icte

d i

nte

stin

al N

dig

esti

on,

Con

dens

ed t

anni

n*

reco

very

(%)

3-w

k le

ngth

13

4.6

99.0

10

8.6

75.0

49

.7

9-w

k le

ngth

97

.4

88.4

87

.6

67.9

48

.1

5.05

'N

= ni

trog

en

'Bir

d-re

sist

ant

sorg

hum

gra

in a

nd b

roile

r li

tter

wer

e th

orou

ghly

mixed,

or b

ird-

resi

stan

t so

rghu

m g

rain

was

pla

ced

abov

e br

oile

r li

tter

in

sepa

rate

laye

rs.

'Per

cent

age

unit

cha

nge

(fin

al m

inus

ini

tial)

. *C

atec

hin

equi

vale

nts.

a a 8 c

h

w.

*c P 4

w

Dow

nloa

ded

by [

Mos

kow

Sta

te U

niv

Bib

liote

] at

13:

44 1

6 Se

ptem

ber

2013

74 Z.S. Wang and wworkrs

responsible. Level of PS did not affect change in soluble true protein concentration. Treatment effects on change in nonprotein N concentration did not occur with 3 wk of deep-stacking. However, at 9 wk change generally increased, from -6.0% for 0% PS to 0.7 and 0.3% for 14.2 and 28.4% PS, respectively. Change in ammonia N concentration with increasing PS level was fairly similar to that with increasing VE level, although the increase in magnitude of ammonia N concentration change was slightly greater for PS than for VE. Effects of level of PS on change in in situ N disappearances and residual pepsin insoluble N after ruminal incubation were not marked but do reflect increasing acid detergent fiber N concentration with increasing PS level.

Change in condensed tannin recovery with increasing PS level (Table 3) was in accordance with that for increasing level of VE. Also, disappearance was greater for 9 us 3 wk of deepatacking. However, condensed tannin disappearance was greater (P<0.05) for mixed than for covered substrates with nonzero PS levels at both 3 and 9 wk; whereas, similar differences for VE substrates with comparable CT levels (1.3, 2.6, 5.1 and 10.3% VE) were noted only at 3 wk. Factors responsible for these effects are not apparent but could involve differences between VE and PS in the nature of condensed tannins or physical charcteristics affecting degree of tannin exposure to conditions in the deep-stack. Alternatively, greater organic matter degradable by microorganisms in PS than may have increased microbial utilization of ammonia, thereby decreasing exposure to condensed tannins.

BS and S Organic matter recovery differed (P<0.05) among BS levels and between lengths of deep-stacking and addition methods, although magnitudes of change were not appreciable (Tables 4 and 5). Organic matter recovery for S substrates increased with increasing S level. Differences in N recovery between deep-stacking for 3 or 9 wk varied with level of BS in substrates, although for S substrates N recovery was greater (P<0.05) for 9 us 3 wk and increased with increasing S level. Magnitudes of change in soluble N concentration were not significantly (BS substrates) or markedly (S substrates) affected by BS or S levels. Change in acid detergent fiber N concentration increased with increases in BS or S level. Thus, these effects did not appear related to condensed tannins in BS. Higher levels of BS and S generally resulted in nonprotein N concentration greater than expected based on concentrations before deep-stacking. Change in ammonia N concentration was greatest (Pc0.05) among BS levels for 40%;

Dow

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by [

Mos

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Sta

te U

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Bib

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] at

13:

44 1

6 Se

ptem

ber

2013

Tab

le 5

E

ffec

ts o

f le

vel o

f re

gula

r so

rghu

m g

rain

and

met

hod

of a

dditi

on t

o br

oile

r li

tter

on

cons

titue

nt r

ecov

ery

and

perc

enta

ge u

nit

chan

ge i

n co

ncen

trat

ion

of c

onst

ituen

ts a

nd in

situ

dis

appe

aran

ce a

fter

dee

p-st

acki

ng f

or

7r 9

wk

Item

' R

egul

ar s

orgh

um grain (9% o

f to

tal

dry

mat

ter)

0

5 10

20

40

SE

3

wk

9w

k

SE

Incu

batio

n le

ngth

Org

anic

mat

ter

reco

very

(%

) Mixed

102.

9 C

over

ed

101.

3 Total

N r

ecov

ery

(%)

92.9

Mix

ed3

1.1

Sol

uble

N c

hang

e (2

')

-1.3

A

cid

det

erg

ent f

iber

N c

hang

e (%

')

cove

red3

0.

5 0.

5 1.

4 In

solu

ble

avai

labl

e N

cha

nge

(%')

Sol

uble

tru

e pro

tein

N c

hang

e (9%')

Am

mon

ia N c

hang

e (%

') N

onam

mon

ia n

onpro

tein

N c

hang

e (%

')

Non

prot

ein

N c

hang

e (9%')

-2.6

3-w

k le

ngth

-3

.7

dis

app

eara

nce

(%')

-0.2

9-w

k le

ngth

-6

.3

Cha

nge

in i

n e

ifu

12-

h N

Cha

nge

in in

eif

u 4

8-h

N

Cha

nge

in p

redic

ted in

test

inal

N

dige

stio

n (9%')

dis

app

eara

nce

(%')

-0.5

3-w

k le

ngth

-0

.2

9-w

k le

n&h

-0.5

102.

9 104.2

104.3

105.

4 10

1.6

102.

2 10

3.6

105.

8 93

.9

94.8

99

.2

99.5

-1

.7

-3.0

-1.0

-0

.3

1.6

1.1

1.7

2.8

0.5

1.9

2.0

5.0

0.7

1.5

-0.9

-3

.6

-1.3

-3

.1

-2.3

-4

.7

-0.4

0.

0 1.

3 4.

4

-1.7

-2

.2

0.1

3.0

-4.3

-4

.3

-3.6

0.

5

-1.5

-3.5

3.5

1.6

Con

dens

ed -

Wn

4 re

cove

ry, 3-wk l

engt

h (%

) M

ked

115.

2 12

0.3

121.

4 13

2.4

133.

5 C

over

ed

108.

0 11

7.5

114.

6 13

2.8

159.

0 'N

= ni

trog

en.

0.28

0.

78

0.60

0.47

0.

66

0.98

0.

71

1.17

1.

33

0.33

0.37

0.55

0.

25

104.

0

94.1

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76 Z.S. Wang and coworkers

whereas, S level did not affect change in ammonia N concentration. Change in nonammonia nonprotein N concentration was inconsistent among BS levels, although nonammonia nonprotein N concentration change increased with increasing S level. Changes in in sitzr disappearances were not markedly or consistently affected by BS or S level in substrates.

Condensed tannin recovery decreased as BS level increased (Table 41, as also was noted for VE and PE substrates. However, BS substrate condensed tannin recoveries were not greatly different from those for VE with levels providing comparable initial substrate condensed tannin concentrations (0.04, 0.09, 0.17 and 0.35% VE). Also in accordance with results for VE and PS substrates, condensed tannin recovery was lower for 9 us 3 wk of deep-stacking. Because of the low level of analyzed condensed tannins in broiler litter and a nondetectable concentration in S, condensed tannin recovery for all S substrates was above 100% and increased slightly with increasing S level.

Summary

The lack of marked effects of condensed tannin source addition on change in concentration of most N fractions in situ disappearances and organic matter and N recoveries indicate no or relatively minor effects on activity of microorganisms in substrate during deep-stacking or of ruminal microbes during in situ incubation. Subjection to broiler litter deep-stack conditions drastically reduced recovery of assayable condensed tannins regardless of source. The decrease in condensed tannin recovery with increasing level of addition indicates that capacity for change elicited by responsible factors (e.g., exposure to ammonia or moist alkaline conditions) was not exceeded. However, it appeared that condensed tannin loss increased with increasing length of deep-stacking and the influence of method of condensed tannin addition differed among sources. With the source highest in condensed tannins (i.e., VE), thoroughly mixing broiler litter with the condensed tannin source resulted in lower condensed tannin recovery than did covering at 3 wk but not at 9 wk; whereas, with the source moderate in condensed tannin concentration (i.e., PS) a difference between addition methods occurred with both lengths of deep- stacking, although it was of lesser magnitude a t 9 wk. Conversely, for the source lowest in condensed tannins (i.e., BS), method of addition did not affect recovery with either deep-stacking length. In conclusion, there appears considerable potential to decrease assayable condensed tannin

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concentration in feedstuffs by addition to broiler litter before deep-stacking.

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