Animal Feed Science and Technology, 9 (1983) 1--17 1
Elsevier Science Publishers B.V., Amsterdam -- Printed in The Netherlands
NUTRI TI VE VALUE FOR RUMI NANTS OF SUGAR CANE BAGASSE ENSI LED AFTER SPRAY TREATMENT WI TH DI FFERENT LEVELS OF NaOH E. M O L I N A , J. B O Z A a n d J.F. A G U I L E R A Estacidn Experimental del Zaidin, Consejo Superior de Investigaciones Cienttficas, Granada (Spain) (Received 2 February 1982; accepted for publ i cat i on 16 December 1982) A B S T R A C T Molina, E., Boza, J. and Aguilera, J.F., 1983. Nutritive value for rumi nant s of sugar cane bagasse ensiled after spray t reat ment with different levels of NaOH. Anim. Feed Sci. Technol., 9: 1--17. Sugar cane bagasse previously dried to a water cont ent below 25% was sprayed with equal quantities of sodium hydroxide solutions to supply 0, 20, 40 or 60 g alkali/kg of dried product and ensiled for 90 days. The cell wall cont ent (NDF) decreased with increasing NaOH (P <~ 0.001), whereas the lignocellulose fraction (ADF) remained unaffected. The effect on lignin (ADL) was small. All this suggests a solubilization of the hemicellulose fraction with alkali applica- tion. Ensfling reduced the pH in the bagasse owing to partial neutralization of residual NaOH. From a microbiological poi nt of view, the stability of the NaOH-treated product, whether during the period of silage or when exposed t o air, was good. The presence of NaOH in the silo inhibited the development of bacteria and fungi, an effect which was probabl y enhanced by the reduced soluble carbohydrate cont ent of the substrate. Sixteen digestibility experiments following a 4 2 2 factorial design were carried out on wethers of 55.0 kg given, ad libitum, isocaloric diets cont ai ni ng the NaOH-spray- ed bagasse suppl ement ed with molasses, a mineral--vitamin mi xt ure and urea--biuret or soya bean meal, to provide a total N cont ent of 1.65 and 2.10%, irrespective of the N source. Average daily intakes were 29.8, 35.7, 44.7 and 40.6 g/kg W 'Ts for diets containing bagasse sprayed with 0, 20, . 40 and 60 g alkali/kg of dried product, respectively. The effects of NaOH level, N source and N level were highly significant (P < 0.001). The apparent digestibility of dry mat t er (DM), organic mat t er (OM) and cell wall component s, and the N ret ent i on increased linearly (P <~ 0.001) as the level of chemical t reat ment increased. Except for hemicellulose, the digestibility of the cell wall compo- nent s was improved when soya bean meal was included in the experimental diets. The effect of N cont ent was not significant. The calculated average organic mat t er digestibili- ty (OMD) for the sugar cane bagasse ranged from 32.6 for the water-treated product to 56.8% with the highest level of alkali, the effect of the chemical t reat ment being sig- nificant (P < 0.001). 0377-8401/ 83/ $03. 00 1983 Elsevier Science Publishers B.V. INTRODUCTION Several aut hors have recent l y reviewed t he alkali t reat ment of pr oduct s rich in lignoceUulose (Owen, 1976; Jackson, 1977; Gonzalez Santillana, 1978; Greenhalgh, 1978; Kl opfenst ei n, 1978; 0rskov, 1979). Sugar cane bagasse (Saccharum officinarum) has a nutritive value l ower t han t hat of ot her l ow-qual i t y roughages such as cereal straws, and t he im- provement obt ai ned by alkali t r eat ment seems t o be inferior (Verma and Jackson, 1975). Ensiling of t he t reat ed pr oduct pr obabl y enhances t he NaOH effect on t he component s of t he cell wails, t hus providing a more available lignocellulose st ruct ure for rumen micro-organisms. The effect of t he i nt ensi t y of alkali spray t r eat ment on t he digestibility in vivo of sugar cane bagasse has not been studied, but t here are dat a from experi ment s in whi ch sugar cane bagasse was t reat ed wi t h alkali according t o Beckman' s t echni que (Randel, 1972) and included in compl et e rations for cattle. On t he ot her hand, it is now clear t hat t he nitrogen requi rement s of t he rumen micro-organisms depend on t he amount of ferment ed subst rat e ( 0r skov et al., 1972; Miller, 1973), and t hat t he nitrogen upt ake is largely a funct i on of bot h t he rate of degradat i on of t he energetic subst rat e and of t he availability of t he nitrogen for micro-organisms (Ol dham et al., 1977). The aim of t he present wor k was t o det ermi ne t he effect of NaOH spray t r eat ment on t he composi t i on and nutritive value for sheep of sugar cane ba- gasse. The ability of a bi ur et - urea mixture, added t o t he experi ment al diets as a non-prot ei n source (NPN), t o meet t he nitrogen requi rement s of rumen micro-organisms was also studied. MATERIALS AND METHODS Experimental procedure Sugar cane bagasse harvested in 1979 f r om t he coast of Granada was dried at ambi ent t emperat ure (spread over a concret e fl oor in t he open air) t o 75% dry mat t er. This partially dried bagasse (several lots of 25 kg per t reat ment ) was spread thinly over a pol yet hyl ene sheet and sprayed wi t h 25 1 of a solu- t i on containing 0, 20, 40 or 60 g NaOH 1-1, at a rate of 2.5 1 mi n -1. Material from each t r eat ment was t hen st ored in t wo silos wi t h a capaci t y of 3 m 3. A represent at i ve sample (5 kg) from each silo was kept anaerobically in a sealed pol yet hyl ene bag for t ot al viable mi crofl ora count s. Isocaloric diets for sheep were prepared by suppl ement i ng t he bagasse wi t h beet molasses, a mi ner al - vi t ami n mi xt ure and a ur ea- bi ur et mi xt ure (60.0% bi uret ; 36.2% urea) (M) or soya bean meal (S), t o provide 1. 65 (I) or 2.10 (II) percent age units of N. The composi t i on of t he diets is shown in Tabl e I. The digestibility trials were carried out on 12-mont h-ol d Segurefia wethers wi t h an average initial weight of 55. 0 kg, individually housed in met abol i sm 3 TABLE I Compos i t i on of di et s cont ai ni ng ur ea- - bi ur et mi xt ur e or s oya be a n meal as the nitrogen s uppl e me nt (g kg -~) Nitrogen sour ce and l evel (%) Ur ea- - bi ur et mi xt ur e Soya bean meal 1. 65 2. 10 1. 65 2. 10 Ensi l ed t r eat ed bagasse ( DM) 650 650 650 650 Beet mol asses 125 125 125 125 Ur ea- - bi ur et mi xt ur e 25 35 - - - - Soya bean meal (8% N) - - - - 125 175 St ar ch 150 140 57 7 Cal ci um t r i phos pha t e 12 12 12 12 Cal ci um di phos pha t e 15 15 15 15 Cal ci um monophos pha t e 3 3 - - - - Magnes i um s ul phat e 14 14 10 10 Magnes i um c a r bona t e 4 4 4 4 Mi ner al - - vi t ami n s uppl e me nt ~ 2 2 2 2 One kg pr ovi des 5 000 000 I U vi t ami n A; 1 600 000 I U vi t ami n D~; 4 g vi t ami n B2; 0. 2 g Co; 0. 75 g I; 1. 2 g Cu; 21 g Zn; 30 g Mn; a nd 42 g Fe. crates and with free access t o water. Sixteen digestibility trials, each on six animals, were per f or med fol l owi ng a 4 (level of alkali) 2 (source of supple- ment al N) 2 (level of di et ary N) factorial design. The intakes were established on t he basis of previous experi ment al dat a obt ai ned on individual animals, t o be t he maxi mum obt ai nabl e wi t hout refusals. When t he ration was not fully consumed, aliquots were t aken for analysis. Aft er a peri od of adapt at i on t o t he diets (25 days when diets were suppl ement ed wi t h t he urea- biuret mi xt ure and 10 days for diets including soya bean meal), t he digestibility and nitrogen balance were det ermi ned in- dividually by collecting t he t ot al excret a for a 10-day peri od, t he animals being fed t wi ce daily. Cumulative samples of feed and excret a were t aken daily for each animal and kept at - 25 C in airtight reservoirs until analysed. Chemical analyses The det ermi nat i ons of dry mat t er, pH and t ot al N were carried out direct- ly on t he samples mai nt ai ned at - 25 C aft er t hawi ng at r oom t emperat ure. The dry mat t er cont ent was det ermi ned by freeze-drying. The pH was also measured in samples of ensiled bagasse 2 and 25 days aft er t reat ment . The analytical measurement s of neutral det ergent fibre (NDF), acid det ergent fibre (ADF), acid det ergent lignin (ADL), ash and silica, and t ot al and resi- dual sodi um, were made on samples oven dried at 60C, equi l i brat ed in air and milled. The NDF was obt ai ned as descri bed by van Soest and Wine (1967), t he ADF and t he ADL according t o van Soest (1963) and silica as t he insoluble mineral fract i on remaining after t r eat ment of ADL ash wi t h 48% HBr for 90 mi n and heating at 5500C. Hemicellulose and cellulose con- t ent s were calculated by di fference ( NDF- ADF and ADF- ADL, respec- tively). The pH det ermi nat i on was carried out wi t h filtered liquid t aken from t he sample aft er macerat i on for 1 h wi t h an equal vol ume of distilled water; t he t ot al N accordi ng t o a macro-Kj eldahl procedure utilizing a mi xt ure of K2SO4, CuSO4 and Se as catalysts; t he mineral cont ent by ashing t he sample at 550C for 3.5 h and t he t ot al sodi um by fl ame phot omet r y aft er minerali- zation at 450C and ext ract i on of t he residue with HC1. The residual sodi um analysis (from NaOH which had not react ed wi t h bagasse) was made by po- t ent i omet ri c evaluation t o pH 8 wi t h 0. 01 N HC1, aft er macerat i on of 10 g of t he sample wi t h 50 ml of distilled water. The organic mat t er digestibility (OMD) of t he bagasse was obt ai ned by assuming t hat t he digestibility coeffi ci ent of t he rest of t he ingredients re- mai ned const ant at 0. 90 in all diets. Microbiological analysis The t ot al viable mi crofl ora was det ermi ned bot h on t reat ed non-ensiled bagasse and on samples st ored anaerobically for 25 days. In t he l at t er case, t he pr oduct stability was st udi ed by exposing samples t o t he open air in Petri dishes for 10 days. In all cases, 10-g samples of bagasse were shaken t horoughl y in 100 ml of distilled water, and 10-fold serial dilutions were prepared f r om t he macerate. Al i quot s (1 ml) of t he sol ut i ons were pour ed in dupl i cat e Petri dishes con- taining a common agar--yeast ext ract medi um with pept one, glucose and salts, and were i ncubat ed at 30C for 5 days. RESULTS Ef f ect o f alkali t reat ment on t he composi t i on o f ensiled sugar cane bagasse The pr et r eat ment mean composi t i on of t he sugar cane bagasse, as well as t he average analytical values of this pr oduct t reat ed wi t h NaOH solutions and ensiled for 90 days, appear in Tabl e II. Over 90% of t he dry mat t er was account ed for by NDF, whi ch consi st ed mai nl y of cellulose (56.0%) and hemicellulose (31.5%), whereas lignin formed onl y 12.5% of this fraction. As expect ed, t he N cont ent was very low. The pH increased consi derabl y wi t h t he i ncorporat i on of 2% NaOH solu- t i on and levelled of f t hereaft er. The pH values in t he t reat ed pr oduct de- creased by about 2.5 units during t he first 25 days in t he silos. An ext ensi on of t he peri od of storage did not i nduce furt her changes (Fig. 1). The cont ent of NDF in t he ensiled sugar cane bagasse decreased (P < 0. 001) T A B L E
I I
E f f e c t
o f
a l k a l i
t r e a t m e n t
a n d
e n s i l i n g
o n
t h e
c o m p o s i t i o n
o f
s u g a r
c a n e
b a g a s s e
( %
d r y
m a t t e r )
P r e - t r e a t e d
L e v e l
o f
N a O H
S E M
'
b a g a s s e
( g / l O 0
g
d r i e d
b a g a s s e )
0
2
4
6
L e v e l
o f
s i g n i f i c a n c e
T r e a t m e n t
L i n e a r
Q u a d r a t i c
e f f e c t
e f f e c t
N D F
9 3 . 6
9 4 . 5
c
8 9 . 5
b
8 6 . 0
b
7 2 . 7
a
1 . 0 2
A D F
6 4 . 1
6 5 . 1
6 4 . 6
6 5 . 2
6 2 . 2
1 . 3 6
A D L
1 1 . 7
1 2 . 0
b
1 1 . 5
b c
9 . 9 3
a c
9 . 0 0
a
0 . 5 5
N D S
6 . 4
5 . 4 7
a
1 0 . 5
b
1 4 . 9
c
2 7 . 3
d
1 . 0 0
H e m i c e l l u l o s e
2 9 . 5
2 9 . 4
c
2 4 . 9
b c
2 0 . 8
b
1 0 . 5
a
1 . 3 1
C e l l u l o s e
5 2 . 5
5 3 . 1
5 3 . 2
5 5 . 3
5 3 . 2
1 . 3 6
T o t a l
N
0 . 5 4
0 . 4 9
0 . 5 2
0 . 4 9
0 . 5 7
0 . 0 3
A s h
3 . 9 9
3 . 2 8
a
7 . 8 5
b
9 . 9 3
b
1 6 . 3
e
0 . 6 3
S i l i c a
1 . 5 0
1 . 5 0
1 . 3 4
1 . 3 4
1 . 5 4
0 . 2 3
T o t a l
N a
- -
T r a c e s
1 . 3 1
2 . 3 2
3 . 7 0
R e s i d u a l
N a
- -
T r a c e s
0 . 3 2
0 . 4 3
1 . 1 7
D r y
m a t t e r
4 1 . 5
4 3 . 9
4 5 . 1
4 5 . 6
4 4 . 2
1 . 6 4
p H
- -
6 . 9 5
9 . 7 0
1 0 . 0
1 0 . 2
N S
- -
- -
* *
* *
N S
* * *
* * *
N S
N S
- -
- -
N S
- -
- -
* * *
* * *
N S
N S
- -
- -
N S
- -
- -
'
S t a n d a r d
e r r o r
o f
m e a n .
a , b , C M e a n s
w i t h i n
a
r o w
w i t h
d i f f e r e n t
s u p e r s c r i p t s
d i f f e r
s i g n i f i c a n t l y ;
P
<
0 . 0 5 .
* * * P <
0 . 0 0 1 ; * * P
<
0 . 0 1 ; * P <
0 . 0 5 ; N S
= P >
0 . 0 5 .
2 d a y s a f t e r e n s i l i n g . . . . . . 25 ,, 1 4 - - . . . . . . . s o / 10" _ _ . . . . - ~ - - . . . . . . --r /- g . N a O H { g / 1 0 0 g d r i e d b a g a s s e ) Fi g. 1. Ef f e c t o f al kal i t r e a t me nt and pe r i od o f ens i l i ng o n pH o f sugar cane bagasse. as t he level of alkali increased, whereas no significant di fferences were f ound in ADF. The fall in NDF cont ent was basically due t o solubilization of hemicellulose (P < 0. 001) and, t o a lesser ext ent , t o t he decrease in t he ADL fract i on (P < 0. 001) caused by t he alkali t reat ment . Significant differences in t he t ot al mineral cont ent of t he pr oduct (P < 0. 001) were found with t he addi t i on of NaOH. Conversely, bot h t he silica and t he N cont ent s of bagasse remained unaffect ed by t he t reat ment s. The results of t he mi crobi al st udy are shown in Tabl e III. Higher count s TABLE IH T o t a l v i a b l e c o u n t o f m i c r o - o r g a n i s m s ( N 1 0 ~ / g f r e s h m a t t e r ) in sugar c a n e bagasse treated w i t h N a O H . P r o d u c t stability after e x p o s u r e t o air NaOH (g/ 100 g dr i ed bagasse) N o n - e n s i l e d bagasse Bact eri a Fungi Ens i l e d bagas s e 0 days I 10 day s ~ Bact eri a Fungi Bact eri a Fungi 0 155 14 - - - - 62 2.0 2 270 30 25 3.0 7. 4 0. 8 4 3 1 1 17 38 0. 5 1. 3 0. 1 6 23 20 18 0. 8 . 1. 7 0. 1 1 Ti me of expos ur e t o air. were f ound in t he non-ensiled product . Ensiling of bagasse for 25 days de- creased 10-fold t he cont ent of viable micro-organisms, irrespective of t he level of alkali t reat ment . The greater levels of alkali also reduced t he number of micro-organisms in non-ensiled bagasse. Aft er its exposure t o air for 10 days t he numbers of bot h t ot al viable bacteria and fungi were substantially reduced. It t hus seemed t hat ensiled alkali-treated sugar cane bagasse was suffi ci ent l y well preserved. Ef f ect of the alkali treatment on intake and digestibility In general, t he daily intakes of t he rations containing 0 or 20 g NaOH/ kg t reat ed bagasse (29. 8 and 35. 7 g/kg W 'Ts, respectively) were t oo l ow and losses of liveweight were observed. Higher alkali levels gave higher intakes (44. 7 and 40. 6 g/kg W 'Ts, respectively, for 40 and 60 g NaOH/ kg t reat ed bagasse), adequat e t o meet mai nt enance requi rement s or t o pr omot e slight gains. The addi t i on of alkali, as well as t he level and source of nitrogen in t he diet, affect ed i nt ake significantly (P < 0. 001). The i nt ake of t he diets containing soya bean meal was superi or t o t hat of rations containing a bi uret --urea mi xt ure. As t he level of NaOH in t he diet increased, t he wat er intake, vol ume of urine and t he wat er cont ent of faeces increased markedl y. The mean digestibility coeffi ci ent s and t he nitrogen ret ent i on results ap- pear in Tabl e IV. Figures 2 and 3 show t he main effect s of t reat ment s. The mean digestibility of diets whi ch included unt reat ed bagasse was 47. 4 and 50.2, respectively, for DM and OM, and rose t o 66. 5 and 68. 0% with t he highest concent rat i on of alkali. Significant linear and quadrat i c component s of variance showed t he i mprovement in digestibility of t he diet as t he level of alkali increased and also t he t endency for t he magni t ude of t he effect t o decline wi t h t he highest level of NaOH addition. The effect s of level and source of di et ary nitrogen were non-significant. Tabl e V gives t he calculated digestibility of organic mat t er (OMD) of t he sugar cane bagasse alone. The average OMD for bagasse ranged from 32. 6 for unt reat ed material t o 56.8% with t he highest level of NaOH addition. Table IV shows t he digestibility coefficients for NDF. The effect of alkali t r eat ment was highly significant; t he linear and quadrat i c component s in- di cat ed an increase in t he digestibility of this fraction t hat was less pro- nounced as t he amount of NaOH in t he bagasse increased. The digestibility of NDF was also significantly affect ed by t he N source but not by t he N level. A qui t e similar pat t ern was found for ADF digestibility. With t he highest level of NaOH in t he diet faecal ADF was higher t han NDF so t he digestibility of hemicellulose was over 100%. Its digestibility was also affect ed by bot h NaOH concent rat i on and N level. The digestibility of t he cellulose was similar t o t hat of hemicellulose, as far as t he NaOH level was concerned. Nevertheless, t wo i mport ant di fferences should be outlined; t he absence of statistical significance for t he effect of di et ary N level and t he high significance for t he N source (P < 0. 01). 8 TABLE I V Mean di gest i bi l i t y coef f i ci ent s and ni t r ogen bal ance i n wet her s f ed on di et s bas ed on ensi l - ed NaOH- t r eat ed sugar cane bagasse (%) T r e a t m e n t D M O M N D F A D F A D L N D S H e m i - Cellulose N re- diet I cellulose tention 0MI 45. 3 47. 4 37. 0 39, 6 25. 1 58. 6 31. 4 42. 6 - - 19. 9 II 51..7 53. 7 42. 5 42, 5 24. 3 66. 0 42. 3 47. 1 - 4. 2 SI 52. 7 56. 4 49. 4 50. 6 30. 0 58. 8 46. 8 54. 2 - - 21. 4 II 41. 3 44. 8 35. 1 31, 2 22. 6 52. 8 43. 4 33. 0 - 14. 9 2MI 56. 4 59. 3 51. 8 46. 3 24. 8 66. 9 47. 2 54. 6 4. 3 II 57. 3 61. 0 54. 7 47. 2 4. 4 60. 6 79. 3 56. 8 5. 0 SI 56. 8 60. 1 53. 4 50. 5 33. 9 62. 5 60. 0 54. 2 0. 1 II 59. 4 62. 5 58. 7 58. 0 40, 9 60. 7 60. 0 61. 6 9. 5 4MI 62. 9 63. 7 61. 7 57. 5 9. 5 65. 8 74. 1 65. 5 28. 8 II 62. 5 65. 2 60. 6 58. 2 31. 6 65. 8 67. 7 63. 3 10. 3 SI 62. 2 64. 6 61. 9 62. 4 35. 7 62. 8 59. 9 67. 4 2. 9 II 63. 1 65. 9 65. 0 65. 0 35. 2 60. 2 65. 0 69. 8 2. 1 6MI 64. 8 65. 6 61. 6 45. 9 - 38. 6 67. 4 130. 0 62. 5 14. 4 II 65. 8 67. 4 66. 9 59. 1 - 5. 9 64. 6 115. 5 69. 7 9. 1 SI 71. 8 74. 5 74. 5 69. 3 2. 1 69. 1 104. 7 78. 3 15. 8 II 63. 6 64. 5 65. 9 52. 5 - 33. 1 61. 5 187. 3 67. 2 10. 8 SE of me a ns 1. 61 1. 60 2. 21 2. 59 5. 26 2. 12 7. 48 2. 05 5. 41 St at i st i cal si gni f i cance of t r e a t me nt s NaOH (1) *** *** *** *** *** *** *** *** *** Li near com- pone nt *** *** *** *** *** *** *** *** *** Quadr at i c c ompone nt *** *** *** *** *** NS *** *** *** Cubi c c ompone nt NS NS NS * *** NS *** NS NS N s our ce (2) NS NS ** *** *** ** NS ** NS N level (3) NS NS NS NS NS * *** NS NS 1 X 2 NS NS NS * * NS * NS * 1 x 3 NS * * ** NS NS * *** *** 2 3 *** *** ** *** ** NS * *** NS 1 2 X 3 *** *** *** *** *** * *** *** ** i 0, 2, 4, 6 = Na OH level; M = ur ea- - bi ur et mi xt ur e , S = s oya bean meal ; I and II = l evel s of s uppl e me nt a t i on. ***P< 0. 001; **P < 0. 01~*P < 0, 05; NS =P> 0. 05. r~ 03 7 0 1 ~ D M . . . . . . O M . . . . N D F x - - x - - x A D F I o - - - o - - o B a g 3 s s e O M 413- 30- / j / / / /1 # // / , / / i / .~ , ' / / I / 1 +1 . : 0 / o / 0 / 0 i / 4- .P'* / / ".1~ + /" j o~ , /o o o/ NaOH( g/ l OOg dried bagasse) Fi g, 2. Ef f e c t o f al kal i t r e at me nt o f sugar cane bagasse on di ge s t i bi l i t y i n v i v o. The di gesti bi l i ty o f the neutral detergent sol ubl es ( SDN) was si gni fi cantl y af f ect ed by NaOH concent rat i on, N l evel and N source. The average values of ADL di gesti bi l i ty f or the di ets wi t h 40 g NaOH/ kg bagasse were ca. 26%. A negati ve di gesti bi l i ty coef f i ci ent was f ound f or t he hi ghest l evel of alkali. Bot h NaOH t reat ment and N source were significant. Ni trogen ret ent i on i mprov ed wi t h the l evel of alkali, but was not af f ect ed by ei ther the N source or l evel . I 0 :5 ~5 60' 50' 60- 50- DM I M OM M S NDF ADF ~ a s s e ) Fig. 3. Ef f ect of l evel and sour ce of di et ar y ni t r ogen on di gest i bi l i t y i n vivo. TABLE V Cal cul at ed di gest i bi l i t y and di gest i bl e or gani c ma t t e r c ont e nt of sugar cane bagasse spray- ed wi t h NaOH s ol ut i ons and ensi l ed (%) Tr e a t me nt I OMD DOM i n DM Level of si gni f i cance' 0MI 27. 4 26. 5 II 37. 7 36. 5 SI 42. 6 41. 2 ( 1) NaOH *** II 25. 1 24. 3 Li near c ompone nt *** Quadr at i c c ompone nt *** 2MI 46. 8 43. 1 Mai n ef f ect s H 46. 1 42. 5 Cubi c c ompone nt NS SI 46. 3 42. 7 ( 2) N sour ce NS II 50. 6 46. 6 ( 3) N l evel NS 4MI 52. 1 46. 9 II 54. 3 48. 9 (1) ( 2) ** SI 53. 8 48. 5 Fi r st or der ( 1) ( 3) NS" II 57. 5 51. 8 i nt er act i ons ( 2) ( 3) *** 6MI 54. 6 45. 7 II 55. 3 46. 3 Second or der SI 67. 2 56. 3 i nt er act i ons II 49. 7 41. 6 SE of means 2. 31 2. 09 ( I ) x ( 2) x ( 3) *** 1 See f oot not e s t o Tabl e IV. II DISCUSSION Our average chemical values for sugar cane bagasse are very close to those of Randel (1972), Sharma (1974) and Marshall and van Horn (1975), and clearly show that its nutritive value is lower than that of cereal straws. The decrease observed in NDF with increasing NaOH has been reported for various roughages, mainly cereal straws, by several workers (Klopfenstein et al., 1972; Gharib et al., 1975; Jackson, 1977; Wilkinson and Gonzalez Santil- lana, 1978) and must be attributable to solubilization of hemicellulose as the ADF fraction is not significantly affected by the alkali t reat ment (Thomsen et al., 1973; Wilkinson and Gonzalez Santillana, 1978). Following the technique of immersion of the material in alkali solution and its subsequent washing, Sharma (1974) obtained orderly increases in the degree of solubilization of sugar cane bagasse upon increasing the alkali con- centration up to 15 g/100 g of bagasse, and no response to higher levels of alkali. A slight reduction in ADL cont ent was found by Ololade et al. (1970), Yu et al. (1975) and Wilkinson and Gonzalez Santillana (1978). The increases in pH and sodium content found with the alkali spray treat- ment are in agreement with those obtained by Fernandez Carmona and Greenhalgh (1972) and Wilkinson and Gonzalez Santillana (1978). The fall of 2.5--3 pH units in the bagasse silages could be due to partial neutraliza- tion of the residual NaOH, both by organic acids formed in the process of ensiling (mainly by residues of uronic acids), and also from hydrolyses of ester linkages of phenyl and acetyl groups bound to xylan chains in the cell wall, which are especially frequent in sugar cane bagasse (Tarkow and Feist, 1969). These reactions seem to be practically completed 25 days after ensil- ing. Our results agree with those of Agrawal (1975), Flipot et al. (1976), Wil- kinson and Gonzalez Santillana (1978) and Fernandez and Gonzalez (1979). Nevertheless, Greenhalgh et al. (1978a) indicate only small reductions in the pH of barley straw treated with 6.6 g of NaOH/100 g of product after being ensiled for I year. The low population of micro-organisms in the ensiled NaOH-treated ba- gasse can be explained by the high pH which affects mainly the fungi, whose optimal pH is below that of bacteria, as well as the inadequacy of the sub- strate for the development of extensive microbial activity. This deleterious effect of NaOH addition on the microflora has been observed by Greenhalgh et al. (1978a, b) and Wilkinson and Gonzalez Santillana (1978) in ensiled cereal straws. The stability of ensiled bagasse after 10 days of exposure to air is rather good, as demonstrated by the decline in total counts of micro-organisms and, particularly, fungi, which are below those found by Wilkinson and Gonzalez Santillana (1978) and Greenhalgh et al. (1978b) in barley straw. The dry mat t er digestibility (DMD) and OMD of diets with untreated ba- gasse were slightly lower than those obtained by Stone et al. (1966) and Randel (1972) in rations containing smaller amounts of bagasse. 12 The OMD of bagasse increased by 24. 2 percent age units aft er spraying wi t h NaOH, whi ch implies an average i mpr ovement of about 4 units per g of alkali/100 g dried bagasse. This is mor e t hen t he i mprovement s descri bed by Fernandez Carmona and Greenhalgh (1972), Greenhalgh et al. (1976), Rexen and Thomsen (1976) and Pirie and Greenhalgh (1978), for diets based on t reat ed cereal straws; by Kategile and Frederi ksen (1979) for diets based on alkali-treated maize cobs and close t o t hose obt ai ned by Kl opfenst ei n et al. (1972). Nevertheless, t he nutritive value of t reat ed sugar cane bagasse was l ower t han t hat of alkali-treated cereal straws, in close agreement wi t h a pre- vious wor k of Verma and Jackson (1975), and similar t o t hat of mor e hgnified roughages aft er alkali t r eat ment (Feist et al., 1970; Guggolz et al., 1971; Choung and McManus, 1976). The ext ent of t he NaOH effect in improving t he digestibility of t he sugar cane bagasse declined as t he NaOH concent rat i on increased; this was possi bl y due t o a rise in t he ruminal osmot i c pressure (Ololade et al., 1972), whi ch coul d reduce t he microbial activity in t he r umen (Bergen, 1970), and t o a faster rate of passage t hrough t he digestive t ract caused by t he increase in wat er intake wi t h t he addi t i on of NaOH (Bol duan et al., 1974; McManus et al., 1976; Berger et al., 1980). Moreover, greater i nt akes were f ound for diets wi t h t he highest levels of NaOH, whi ch woul d reduce t he ret ent i on of digesta in t he rumen and, t herefore, t he length of t i me for mi crobi al activity. The i mprovement in OMD of t reat ed bagasse is due t o an increase in t he digestibility of t he cell wall component s (except lignin), derived from bot h their partial solubilization and t he greater availability of t he non-solubilized material for mi crobi al breakdown. The increase in t he digestibility of hemi- cellulose is pr obabl y due t o t he release of phenyl and acet yl groups, especial- ly abundant in t he Graminae (Morris and Bacon, 1976). Lignin artifacts soluble in neut ral det ergent but insoluble in acid det ergent appear in faeces wi t h t he higher levels of alkali and i mpl y t hat t he faecal ADF is higher t han t he NDF fraction; thus, digestibility coeffi ci ent s for hemicellulose above 100% are obt ai ned. Rexen and Thomsen (1976) f ound similar results with barl ey straw. These compounds similar t o lignin, col l ect ed in t he faecal ADF (Hart l ey and Jones, 1978), coul d explain t he negative digestibility coeffi- cients of ADL f ound wi t h t he highest levels of alkali. Allison and Osbourn (1970) and Fahey et al. ( 1979) obt ai ned negative digestibility coeffi ci ent s even l ower t han t hose r epor t ed in this paper. The increase in t he digestibility of cellulose after alkali t r eat ment is attri- but abl e t o bot h t he breakage of hydrogen bonds (Whistler and Teng, 1970; Bacon, 1979) and t he saponi fi cat i on of ester linkages bet ween cellulose and hemicellulose mol ecul es (Feist et al., 1970). The ADL digestibility coeffi ci ent s ranged bet ween 25 and 28% for t he bagasse t reat ed wi t h 0--40 g NaOH/ kg; values greater t han t hose for gross sugar cane bagasse found by Fahey et ah (1979). It seems t hat in t he presence of molasses and starch, sol ubl e l i gni n- car bohydr at e compl exes are f or med by mi crobi al act i on in t he rumen, whi ch are not recovered in t he acid deter- 13 gent residue (Gaillard and Richards, 1975). This would explain the partial digestibility of lignin, an effect found also by Allison and Osbourn (1970), Minson (1971) and Grant et al. (1974), which raises doubts on the suit- ability of lignin as an indicator for digestibility trials. In our experiments the microbial growth was probably more limited by the rate of degradation of the energetic substrate than by the availability of N, and thus the N level did not affect either the DMD, OMD, cell wall digestibility (with the exception of hemicellulose) or the nitrogen retention. Our results are in disagreement with those of Donefer et al. (1969), (prskov and Grubb (1978) and Kategile (1979), who found a higher digestibility of the diet after the addition of NPN. The source of supplemental nitrogen affected the digestibility of cell wall components (except hemicellulose), which was higher with soya bean meal, suggesting more efficient utilization of soya protein than of NPN by the ruminal microflora. This low efficiency of utilization of NPN can also be the . ,o l - - DDM i n v i t r o . . . . . . . DOM . . . . . . . . DDM i n s a c c o x - - x - - x DOM . . . . o - - o ~ o DNDF . . . . ~--A--A#ADF - " s Xi j / I t / * / / < + .:6" /Z I # 0~0~ "O~D o , ' o 2'0 ~ o ~ o i o O i s g e s t i b i l i t y i n v i t r o o r i n s a c c o ( %) Fig. 4. Di gest i bi l i t y in v i tro or i n sacco of NaOH- t r eat ed bagasse as pr edi ct or s of digest- i bi l i ty i n vivo. 14 result of an incomplete adaptation of the ruminal microflora to biuret. Schroder (1970) found that up to 70 days were necessary in order to attain the maximum biuretolytic activity in the rumen, depending on the protein cont ent in the supplemented ration. With regard t o the effect of alkali treatment, the N balance showed the same pattern as the DMD and OMD. We have obtained similar results to those found by Klopfenstein et al. (1972) and Donaldson et al. (1976), who reported better N retention with NaOH treatment. It follows that the struc- ture of the bagasse prevents adequate microbial growth, which is limited by the availability of energy. This fact would explain the negative nitrogen balances reported (Qrskov and Grubb, 1978; (~rskov, 1979). The appearance of significant second- and third-order interactions is diffi- cult to explain and restrains the magnitude of the effect t o our experimental conditions. Figure 4 shows the relationship between digestibility of organic matter in vivo and in vitro or in sacco, previously determined for different fractions in samples of the alkali-treated sugar cane bagasse. 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