Acidogenic value of feeds. I.

The relationship between the

acidogenic value of feeds and in vitro ruminal pH changes
B. Rustomo
1
, J. P. Cant
1
, M. Z. Fan
1
, T. F. Duffield
2
, N. E. Odongo
1
, and B. W. McBride
1,3
1
Department of Animal and Poultry Science,
2
Department of Population Medicine, University of Guelph, Guelph,
Ontario, Canada N1G 2W1. Received 21 October 2004, accepted 28 December 2005.
Rustomo, B., Cant, J. P., Fan, M. Z., Duffield, T. F., Odongo, N. E. and McBride, B. W. 2006. Acidogenic value of feeds. I. The
relationship between the acidogenic value of feeds and in vitro ruminal pH changes. Can. J. Anim. Sci. 86: 109117. The objec-
tive of this study was to use an in vitro technique (i) to assess the acidogenic value (AV) of feed ingredients, (ii) to evaluate the rela-
tionship between the AV of feed and ruminal pH changes, and (iii) to determine the relationship between the AV of feeds and
chemical constituents of feeds. Assessments of AV were based on 24 and 48 h in vitro incubation in rumen liquor. A series of feeds,
ranging from energy, fibre and protein sources were evaluated. Ruminal fluid pH changes in the incubation medium were measured
at the end of 24 and 48 h incubation in buffered rumen liquor (60% buffer, 40% rumen liquor). The chemical constituents of the feed
ingredients were determined using standard procedures. There were no differences (P > 0.05) between 24 and 48 h incubations on
apparent AV and rumen fluid pH changes. The best predictors of AV for all classes of feed were non-fibre carbohydrate (NFC) frac-
tion and acid detergent fibre (ADF; R
2
= 0.81; P < 0.001). The best predictor of AV for energy sources were NFC and ADF (R
2
=
0.70; P < 0.027); neutral detergent fibre (NDF) for fibre sources (R
2
= 0.84; P < 0.027) and crude protein (CP) for protein sources
(R
2
= 0.73; P < 0.014). The rumen fluid pH changes had stronger relationships with apparent AV of all feeds after 24 h (R
2
= 0.74, P
< 0.0001) than starch (R
2
= 0.35, P = 0.04) or NFC (R
2
= 0.56; P < 0.0001). The results indicate that 24 h AV measurements and
rumen fluid pH changes are acceptable measures for qualitatively describing or ranking feed ingredients.
Key words: Acidogenic value, dairy cow, feed chemical composition, rumen pH
Rustomo, B., Cant, J. P., Fan, M. Z., Duffield, T. F., Odongo, N. E. et McBride, B. W. 2006. Valeur acidogne des aliments du
btail. I. Relations entre la valeur acidogne des aliments du btail et la variation in vitro du pH du rumen. Can. J. Anim.
Sci. 86: 109117. Par le biais dune technique in vitro, ltude devait : (i) valuer la valeur acidogne (VA) des aliments du btail,
(ii) tablir les liens entre la VA des aliments du btail et les changements de pH dans le rumen et (iii) prciser la relation entre la
VA des aliments du btail et leur composition chimique. La VA a t value par incubation in vitro dans du fluide du rumen pen-
dant 24 h ou 48 h. Les auteurs ont examin plusieurs types daliments reprsentant diverses sources dnergie, de fibres et de pro-
tines. Ils ont mesur lcart du pH du fluide du rumen dans le milieu dincubation au terme de la priode dincubation de 24 h ou
de 48 h dans du fluide du rumen tamponn (60 % de solution tampon, 40 % de fluide du rumen). La composition chimique des
aliments du btail a t tablie de la manire usuelle. La VA apparente et la variation de pH du fluide du rumen sont les mmes
(P > 0,05) aprs 24 ou 48 h dincubation. La fraction dhydrates de carbone non cellulosiques (FNC) et les fibres au dtergent
acide (FDA) prdisent le mieux la VA, peu importe le type daliment (R
2
= 0,81, P < 0,001). La FNC et les FDA prdisent le mieux
la VA pour les sources dnergie (R
2
= 0,70, P < 0,027); pour les sources de fibres, il sagit des fibres au dtergent neutre
(R
2
= 0,84, P < 0,027) tandis que pour les sources de protines, la protine brute donne les meilleurs rsultats (R
2
= 0,73,
P < 0,014). La variation de pH du fluide du rumen prsente des liens plus troits avec la VA apparente des aliments du btail aprs
24 h (R
2
= 0,74, P < 0,0001) que lamidon (R
2
= 0,35, P = 0,04) ou la FNC (R
2
= 0,56, P < 0,0001). Les rsultats indiquent quon
pourrait recourir la quantification de la VA aprs 24 h dincubation et la variation du pH dans le fluide du rumen pour dcrire
ou classer les aliments du btail qualitativement.
Mots cls: Valeur acidogne, vaches laitires, composition chimique des aliments, pH du rumen
Sub-acute rumen acidosis (SARA) represents lowered rumi-
nal pH as a result of carbohydrate fermentation (Nocek
1997). Effective fibre intake and carbohydrate digestion rate
interact to determine ruminal pH (Armentano and Pereira
1997). Rumen acidosis is therefore related to the amount of
acid produced as feed is fermented and the ability of the feed
to encourage chewing and production of salivary buffers.
Although animals suffering from SARA do not typically
exhibit clinical signs of illness and often go undetected
(Owens et al. 1998), SARA can reduce feed intake, nega-
tively affect rumen fermentation, growth, performance, and
contribute to laminitis (Nocek 1997). Sub-acute ruminal aci-
dosis can also damage ruminal and intestinal epithelial tis-
sue, leading to bacterial infection and subsequent liver
abscesses (Underwood 1992).
Wadhwa et al. (2001) have developed a simple laboratory-
based technique for evaluating acid production from feedstuffs
109
3
To whom correspondence should be addressed (e-mail:
bmcbride@uoguelph.ca).
Abbreviations: AV, acidogenic value; ADF, acid deter-
gent fibre; BC, buffering capacity; CP, crude protein; DMI,
dry matter intake; NDF, neutral detergent fibre; NFC, non-
fibre carbohydrate; NPN, non-protein nitrogen; SARA, sub-
acute rumen acidosis
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110 CANADIAN JOURNAL OF ANIMAL SCIENCE
based on the dissolution of Ca from CaCO
3
. This method has
been used to rank feeds in terms of the acid-load accumulated
within the rumen during fermentation. The concept of effec-
tive neutral detergent fibre (eNDF), which integrates the
effects of diet on chewing, saliva production, ruminal acid pro-
duction and neutralization, uses milk fat composition as a
response variable to predict acidosis (Armentano and Pereira
1997). However, Pereira et al. (1999) showed that milk fat
depression was not a reliable indicator of peNDF or SARA.
The concept of physically effective NDF (peNDF) to describe
the impact of physical effectiveness of NDF in stimulating cud
chewing has also been described (Mertens 1997). Although
the concept of peNDF was developed to avoid the problem of
excess production of acid in the rumen (Mertens 1997), this
approach does not consider acid production from the feed.
Other in vitro studies have determined acid production from
feeds by measuring the final pH after incubation (de Smet et
al. 1995; Malestein et al. 1982). However, pH measurements
alone do not include aspects of buffering (Stewart 1983). In
the acidogenic value (AV) approach, the dissolved Ca repre-
sents how much total acid is produced and neutralized during
fermentation (Wadhwa et al. 2001).
However, Wadhwa et al. (2001) did not measure the rela-
tionship between AV and ruminal fluid pH changes in the
incubation medium for different classes of feeds.
Additionally, Wadhwa et al. (2001) did not measure the
relationship between AV and the chemical components
from different classes of feeds. The objectives of this study,
therefore, were: (i) to measure the AV of feeds, (ii) to eval-
uate the relationship between AV of feeds and ruminal pH
changes, and (iii) to examine the relationship between AV
or ruminal fluid pH changes and feed chemical composition.
MATERIALS AND METHODS
The AV of the feed was determined using an in vitro technique
developed by Tilley and Terry (1963) and modified by
Wadhwa et al. (2001). Feeds were freeze-dried and ground
through a 1-mm screen in a laboratory mill (Thomas Wiley,
Philadelphia, PA) before being returned to the freeze drier to
remove any moisture that had been picked up during grinding.
One-gram (DM basis) samples were weighed directly from the
freeze drier into 100-mL incubation tubes held at 39C in a
water bath. The samples were incubated in duplicate with 30
mL of buffered rumen liquor comprising 60% buffer and 40%
rumen liquor. The buffer (5.880 g L
1
NaHCO
3
; 5.580 g L
1
Na
2
HPO
4
; 0.282 g L
1
NaCl; 0.342 g L
1
KCl; 0.028 g L
1
CaCl
2
.2H
2
O and 0.036 g L
1
MgCl
2
) was made up at 20% the
strength of the Tilley and Terry (1963) buffer (Wadhwa et al.
2001). Rumen fluid was collected from two rumen-fistulated
cows fed alfalfa hay ad libitum 3 h after morning feeding. All
experimental procedures using the fistulated cows were done
with the approval of the University of Guelph Animal Care
Committee in accordance with the guidelines of the Canadian
Council on Animal Care.
Cysteine hydrochloride monohydrate (0.025% wt/vol)
was added into the 100-mL incubation tubes just before
incubation and the tubes were closed with gas release valves
and shaken continuously. After 24 and 48 h of incubation, 2-
mL samples were withdrawn from each tube and transferred
in to 8-mL centrifuge tubes containing 50 mg of CaCO
3
powder (Catalogue No. C6763; Sigma Chemical Co., St.
Louis, MO). The mixture was shaken manually for 5 s and
then centrifuged at 4000 g for 10 min and the Ca content
in the supernatant determined using a test kit (Sigma
Diagnostics Inc., Calcium Procedure No. 587; Sigma-
Aldrich Co., St. Louis, MO) in a laboratory spectropho-
tometer (HACH, DR/4000, Loveland, CO) set at 575 nm.
The absorbance was read to zero using water as reference.
All samples were run in duplicates in two separate runs.
Apparent AV was calculated as the product of Ca concen-
tration from the analysis and fluid volume (30 mL) divided by
the sample weight (1 g; Wadhwa et al. 2001). To eliminate the
contribution of Ca from the feed, basal AV was calculated
after correcting for Ca dissolved before the addition of CaCO
3
.
True AV was calculated as apparent AV (after CaCO
3
) less
basal AV (before CaCO
3
). Blank (with no feed sample) sam-
ples and standards (wheat and straw) were included in each
run for calibration but these were not used to adjust the AV.
The pH was measured before (0 h) and after incubations (24
and 48 h). Feed ingredients chosen for assessment included a
range of energy, protein and fibre sources.
The chemical composition of the feed ingredients was
determined using standard Association of Official
Analytical Chemists [(AOAC) 1990] procedures. The DM
content was determined by oven drying at 60
o
C for 48 h.
Crude protein (CP), soluble protein and non-protein nitro-
gen (NPN) were analyzed using macro-Kjeldahl (method
984.13). Starch using IKA analyzer technick (C5000) and
Ca and P by inductively coupled plasma spectroscopy
(method 945.46). The samples were also analysed for fat,
non-fibre carbohydrate (NFC), acid detergent fibre (ADF;
method 973.18c) and lignin (AOAC 1990) and neutral
detergent fibre (NDF) (Goering and Van Soest 1970).
Statistical Analysis
The relationship between AV and rumen fluid pH and AV and
feed chemical composition was determined using simple and
multiple linear regression analysis within the SAS Institute,
Inc. (2004). Comparison between 24 and 48 h incubations
were conducted using paired t-test procedures in SAS. Effects
were considered significant at a probability P < 0.05.
RESULTS AND DISCUSSION
The chemical composition of the feed ingredients is present-
ed in Table 1. Acidogenic values for feed ingredients after 24
and 48 h of incubation are shown in Table 2. The basal AV of
some feed ingredients (e.g. sugar beet pulp and alfalfa hay)
were relatively high (6.15 0.29 and 5.44 0.21, respective-
ly; Table 2), which suggests contribution of inherent Ca with-
in these feed ingredients to the measured apparent AV.
Energy sources had the highest AV; fibre sources had inter-
mediate AV and protein sources had the lowest AV.
Acidogenic values were not different (P > 0.05) between the
24 and 48 h incubation for all feed classes (Table 3). This sug-
gests that there was little further fermentation after 24 h. It has
been shown that the rate of degradation of feed ingredients is
higher during the first 9 to12 h than during subsequent incu-
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RUSTOMO ET AL. ACIDOGENIC VALUE OF FEEDS. I 111
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112 CANADIAN JOURNAL OF ANIMAL SCIENCE
bation (de Smet et al. 1995). The differences in AV were like-
ly related to the rapid initial fermentation of the feeds. de
Smet et al. (1995) also showed that the rate of degradation
varied with the type of feed ingredient, being higher for feeds
with high cell content and lower for feeds with high cell wall
content. Energy source feeds resulted in similar AV with fibre
sources (Table 2) but had the greatest rumen fluid pH changes
(Table 3 and Fig. 1a).
Table 2. Acidogenic value [AV; dissolved Ca (mg Ca g
1
feed DM)] of feed ingredients after 24 and 48 h incubations. Values are given for Ca dissolved
before (basal AV) and after the addition of CaCO
3
(apparent AV) and the difference (true AV). Standard error of the mean is given in parentheses
24 h 48 h
Feed ingredients Apparent AV Basal AV True AV Apparent AV Basal AV True AV
Energy sources
Sugar beet pulp 14.68 (0.43) 6.15 (0.29) 8.53 (0.66) 13.14 (0.34) 5.37 (0.06) 7.77 (0.38)
Barley 12.67 (0.36) 3.60 (0.34) 9.06 (0.54) 11.28 (0.11) 3.07 (0.06) 8.21 (0.09)
Oats 12.41 (0.22) 2.98 (0.21) 9.43 (0.41) 11.13 (0.27) 2.91 (0.12) 8.22 (0.16)
Wheat 11.65 (0.62) 3.28 (0.28) 8.37 (0.87) 11.06 (0.12) 2.74 (0.13) 8.32 (0.21)
Wheat middling 10.84 (0.94) 3.52 (0.23) 7.31 (0.17) 10.56 (0.37) 3.92 (0.19) 6.64 (0.54)
High moisture corn 9.59 (0.24) 2.14 (0.23) 7.44 (0.31) 11.53 (0.25) 2.43 (0.22) 9.10 (0.45)
Corn distillers 8.38 (0.22) 2.67 (0.30) 5.71 (0.29) 9.26 (0.17) 2.58 (0.20) 6.68 (0.24)
Wheat shorts 8.04 (0.81) 3.17 (0.34) 4.87 (0.78) 9.67 (0.41) 2.97 (0.11) 6.70 (0.31)
Wheat bran 5.70 (0.19) 2.84 (0.27) 2.86 (0.26) 8.35 (0.17) 2.70 (0.18) 5.64 (0.20)
Fibre sources
Corn silage 12.86 (0.25) 3.60 (0.22) 9.24 (0.29) 12.20 (0.18) 3.73 (0.11) 8.46 (0.11)
Alfalfa pellet 12.63 (0.67) 4.92 (0.42) 7.71 (0.26) 10.41 (0.28) 4.01 (0.39) 6.41 (0.25)
Alfalfa hay 11.71 (0.66) 5.44 (0.21) 6.09 (0.50) 11.46 (0.15) 4.27 (0.31) 7.18 (0.31)
Haylage 9.30 (0.50) 5.28 (0.34) 4.02 (0.28) 7.95 (0.36) 3.86 (0.17) 4.09 (0.21)
Wheat straw 6.27 (0.90) 3.04 (0.27) 3.22 (0.64) 6.78 (0.56) 2.69 (0.17) 4.09 (0.40)
Protein sources
Roasted soybean 7.62 (0.68) 2.34 (0.34) 5.28 (0.39) 5.45 (0.32) 1.57 (0.18) 3.88 (0.17)
Soybean meal 6.43 (0.39) 3.54 (0.33) 2.89 (0.39) 5.48 (0.40) 2.69 (0.08) 2.78 (0.37)
Canola meal 5.13 (0.43) 3.65 (0.13) 1.48 (0.32) 4.09 (0.16) 2.54 (0.30) 1.55 (0.46)
Feather meal 3.03 (0.20) 0.79 (0.08) 2.24 (0.17) 1.40 (0.18) 0.34 (0.03) 1.06 (0.16)
Corn gluten 2.40 (0.24) 2.13 (0.30) 0.27 (0.21) 3.10 (0.16) 2.07 (0.28) 1.02 (0.34)
Blood meal 1.66 (0.11) 1.33 (0.12) 0.33 (0.16) 1.31 (0.19) 0.57 (0.05) 0.74 (0.15)
Herring meal 1.42 (0.22) 0.97 (0.09) 0.46 (0.28) 1.87 (0.24) 0.63 (0.07) 1.24 (0.29)
Table 3. Effects of incubation time on basal, apparent and true acidogenic value [AV; dissolved Ca (mg Ca g
1
feed DM)] and rumen fluid pH changes
during 24 and 48 h incubations
24 h 48 h SEM
z
P value
All feed classes
Apparent AV (after the addition of CaCO
3
) 8.30 7.97 0.31 0.461
Basal AV (before the addition of CaCO
3
) 3.21 2.75 0.11 0.003
True AV
y
5.09 5.23 0.24 0.680
Rumen fluid pH changes
x
1.30 1.39 0.07 0.478
Energy sources
Apparent AV (after the addition of CaCO
3
) 10.44 10.66 0.27 0.570
Basal AV (before the addition of CaCO
3
) 3.37 3.19 0.13 0.338
True AV
y
7.07 7.47 0.24 0.250
Rumen fluid pH changes
x
1.75 1.90 0.05 0.107
Fibre sources
Apparent AV (after the addition of CaCO
3
) 10.52 9.76 0.42 0.207
Basal AV (before the addition of CaCO
3
) 4.46 3.71 0.17 0.003
True AV
y
6.06 6.05 0.35 0.986
Rumen fluid pH changes
x
1.36 1.53 0.07 0.199
Protein sources
Apparent AV (after the addition of CaCO
3
) 3.96 3.24 0.27 0.086
Basal AV (before the addition of CaCO
3
) 2.11 1.49 0.15 0.005
True AV
y
1.85 1.75 0.22 0.757
Rumen fluid pH changes
x
0.68 0.65 0.09 0.884
z
Standard error of the mean.
y
True AV was calculated as apparent AV (AV after addition of CaCO
3
) basal AV (AV before addition of CaCO
3
).
x
Rumen fluid pH changes = decrease in rumen fluid pH from the start of incubation to the end of incubation.
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RUSTOMO ET AL. ACIDOGENIC VALUE OF FEEDS. I 113
Fig. 1. (a) Rumen fluid pH changes from 0 h to 24 and 48 h after incubation for the energy sources. (b) Rumen fluid pH changes from
0 h to 24 and 48 h after incubation for the fibre sources. (c) Rumen fluid pH changes from 0 to 24 h and 48 h after incubation for the pro-
tein sources.
(a)
(c)
(b)
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114 CANADIAN JOURNAL OF ANIMAL SCIENCE
Rumen fluid pH changes for different classes of feed
ingredients after 24 and 48 h of incubation are shown in
Fig. 1. The rumen fluid pH decreased significantly during
the first 24 h of incubation, but remained relatively constant
after that to 48 h of incubation (Fig. 1 a, b). The rate of
change of rumen fluid pH changes appears to follow similar
patterns as the AV changes. Energy source feed ingredients
showed the greatest pH decrease during the first 24 h of
incubation (Fig. 1a). This result is in agreement with those
of de Smet et al. (1995) and Hall (2002), that highly fer-
mentable carbohydrates, such as sugars, soluble fibre and
some starches, have the capacity to decrease ruminal pH in
a relatively short period of time (1 to 5 h). Rumen fluid pH
changes were lowest for protein feed sources (Fig. 1c). The
differences in rumen fluid pH changes between 24 and 48 h
of incubation were also smallest for protein feeds (Table 3).
These results suggest that protein feed sources have differ-
ent effects on ruminal pH changes. It has been shown that
high protein feeds generally have higher buffering capacity
(McBurney et al. 1983; Jasaitis et al. 1987) due to the
ammonia produced during fermentation (Crawford et al.
1983). High protein feeds are therefore capable of neutraliz-
ing acid to maintain constant pH in continuous culture
(Dewhurst et al. 2001).
The relationship between apparent AV and rumen fluid pH
changes after 24 h of incubation are presented in Table 4.
There was a positive correlation between AV and rumen fluid
pH change for all classes of feed (R
2
= 0.74; P < 0.0001;
Table 4). High AV was associated with a greater decrease in
rumen fluid pH change. Furthermore, the significant relation-
ship between apparent AV and rumen fluid pH changes sug-
gests high AV diets would be expected to increase the risk of
rumen acidosis in cows. Several studies have shown
decreased ruminal pH when more rapidly fermentable carbo-
hydrates were included in the diet (Keunen et al. 2002;
Krause et al. 2002b; Plaizier et al. 2001; Reinhart et al.1993).
It has also been suggested that increased fermentation of
starch might overload the absorptive capacity of the rumen
thus exacerbating the reductions in ruminal pH (Krause et al.
2003). Furthermore, a decrease in ruminal pH was reported to
decrease appetite (Briton and Stock 1987), fibre degradabili-
ty (Hoover 1986; Krajcarski-Hunt et al. 2002), microbial pro-
tein synthesis (Strobel and Russell 1986; de Veth and Kolver
2001; Calsamiglia et al. 2002), DMI and subsequent milk pro-
duction (Aldrich et al. 1993; Krause et al. 2002a). Prediction
of potential rumen fluid pH changes based on the AV of feeds
would be highly beneficial with regard to diet formulation in
order to balance for diets that would optimize ruminal pH and
rumen function. However, in vivo studies are needed to clar-
ify the effect of feed AV on actual ruminal pH changes in
dairy cows.
The relationships between the AV of the feeds and feed
chemical composition are presented in Table 5. There was a
positive correlation between AV and starch content (R
2
= 0.20;
P = 0.042) of feed and AV and NFC content (R
2
= 0.43; P <
0.001) of the feed. Although the starch contents of sugar beet
pulp and corn distillers were low, their NFC contents were
high (Table 1). Sugar beet pulp contains a significant amount
of soluble fibre (17.4 to 30.0% DM), and sugars (12.8 to
24.7% DM), while corn distillers contain soluble fibre in the
range of 7.8 to 11.6% DM, and sugars from 3.2 to 14.5% DM
(Hall et al. 1999). Sugars and starch ferment to lactic acid,
which has a lower pKa than the volatile fatty acids. Soluble
fibre, such as pectin, ferments rapidly, but its rate of fermenta-
tion declines at low pH (Ben-Ghedalia et al. 1989; Strobel and
Russel 1986). Although the starch and NFC content of high-
moisture corn were the highest among the energy feeds (Table
1), the AV was intermediate (Table 2). Different types of NFC
differ in their rate of fermentation and their effects on ruminal
pH (de Smet et al. 1995; Hall 2000). Rooeny and Pflugfelder
(1986) reported that the starch granules in corn are almost
completely contained within a protein matrix, which decreas-
es the availability of starch to hydrolysis. For all classes of
feed, the relationship between rumen fluid pH changes and
AV were stronger (R
2
= 0.74; P < 0.001) than the relationship
between starch and rumen fluid pH changes (R
2
= 0.35; P =
0.004) or rumen fluid pH changes and NFC (R
2
= 0.56; P <
0.001; Table 4). Crude protein content was negatively corre-
lated with AV (Table 5). High protein feeds would therefore
be expected to have low fermentability and thus produce less
rumen acid load. As noted above, it has been shown that high
protein feeds generally have higher buffering capacity
(McBurney et al. 1983; Jasaitis et al. 1987) due to the ammo-
nia produced during fermentation (Crawford et al. 1983). It is
therefore, important to consider feed ingredient AV when for-
mulating rations rather than just using the starch content or the
NFC content.
Among the forages tested, corn silage had the highest AV
and wheat straw the lowest (Table 2). Corn silage was
almost as acidogenic as the high-energy feeds despite its
lower starch content and higher NDF content (Table 1). The
high AV for corn silage might suggest high levels of free
acids (Thomas and Wilkinson 1975) and low CP content
(Table 1). Additionally, it has been shown that corn silage
required higher alkali to stabilize high rumen fluid pH
changes in continuous culture (Crawford 1983; Dewhurst et
al. 2001; Wadhwa et al. 2001). The low AV for wheat straw
might be explained by its poor NDF degradability.
According to Weiss et al. (1989) and de Smet et al. (1995),
feedstuffs rich in cell wall content have low degradation
rates. These results support the use of an optimum forage
mixture when feeding large amounts of starch-rich concen-
trates to dairy cows (Phipps et al. 1995).
Table 6 shows apparent AV predictions from chemical
composition of the dietary ingredients based on stepwise
multiple linear regression analysis. Greater variation was
explained when the factors were included in combination
demonstrating the importance of the interactions among
chemical constituents in a diet. This suggests that acid pro-
duction and neutralization in the rumen is a complex phe-
nomenon and involves the interaction between feed
components, ruminal environment and microbial popula-
tions. More species of microorganisms grow in the rumen
when the substrates consist of a more complex composition
(Malestein et al. 1982). de Smet et al. (1995) postulated that
a synergism between different bacterial species may occur
when a substrate is more complex and fermentation prod-
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RUSTOMO ET AL. ACIDOGENIC VALUE OF FEEDS. I 115
ucts of one species are used as substrate by another species.
However, some feed ingredients have a poor balance of sub-
strates for rumen microbes. Wadhwa et al. (2001) found that
some feed ingredients such as wheat (high starch content),
wheat straw (high fibre), and feather meal (high protein con-
tent) had deviations from a linear response on AV.
Furthermore, the extent of ruminal fermentation for both
fibre and non-fibre carbohydrates among feedstuffs is
extremely variable and is influenced by interactions
between diet composition and rumen microbial activity
(Allen 1997). Measures of feed chemical composition alone
are therefore inadequate for estimating acid-load in the
rumen and may not be useful as AV indicators.
CONCLUSIONS
This study shows that the acid loads from feeds can be esti-
mated by an in vitro technique. Energy sources and fibre
sources had the highest AV whilst protein sources had the
lowest AV. There was a positive correlation (P < 0.001)
between AV and rumen fluid pH changes for all feeds. A high
AV was associated with a greater decrease in rumen fluid pH
change. The rumen fluid pH changes had stronger relation-
ship with apparent AV of all feeds after 24 h (R
2
= 0.74, P <
0.0001) than with starch (R
2
= 0.35, P = 0.04) or with NFC
(R
2
= 0.56; P < 0.0001). This relationship may allow predic-
tions of ruminal fluid pH change from feed AV. The AV esti-
mates of a range of feed ingredients can be used to formulate
Table 4. Prediction
z
of rumen fluid pH changes
y
(Y) from apparent acidogenic value [AV; dissolved Ca (mg Ca g
1
feed DM)] of feed ingredients (x)
after 24 h of incubation
Factor (x) b
0
b
1
b
2
b
3
R
2
P value
All feeds
Y
1
= b
0
+ b
1
x
1
(starch ) 1.046 0.014 0.35 0.004
Y
2
= b
0
+ b
2
x
2
(NFC
x
) 0.795 0.018 0.56 <0.0001
Y
3
= b
0
+ b
3
x
3
(AV) 0.313 0.119 0.74 <0.0001
Energy sources
Y
1
= b
0
+ b
1
x
1
(starch ) 1.71 0.001 0.02 0.72
Y
2
= b
0
+ b
2
x
2
(NFC) 1.57 0.004 0.17 0.26
Y
3
= b
0
+ b
3
x
3
(AV) 1.19 0.054 0.47 0.04
Fibre sources
Y
1
= b
0
+ b
1
x
1
(starch ) 1.23 0.015 0.61 0.118
Y
2
= b
0
+ b
2
x
2
(NFC) 0.99 0.018 0.62 0.114
Y
3
= b
0
+ b
3
x
3
(AV) 0.42 0.089 0.80 0.040
Protein sources
Y
1
= b
0
+ b
1
x
1
(starch ) 0.59 0.037 0.17 0.351
Y
2
= b
0
+ b
2
x
2
(NFC) 0.29 0.036 0.82 0.005
Y
3
= b
0
+ b
3
x
3
(AV) 0.17 0.130 0.58 0.048
z
Rumen fluid pH changes after 24 h incubations (Y
1
) = b
0
+ b
1
(x
1
); (Y
2
) = b
0
+ b
2
(x
2
); where b
0
= intercept, b
1
= slope (increase in pH changes per unit
increase in starch); b
2
= slope (increase in pH changes per unit increase in NFC); x
1
= starch content and x
2
= NFC content ; and b
3
= slope (increase in pH
changes per unit increase in apparent AV); x
1
= starch content; x
2
= NFC content, and x
3
= apparent AV.
y
Rumen fluid pH change = rumen fluid pH at the start of incubation rumen fluid pH at the end of 24 h of incubation.
x
NFC = non-fibre carbohydrate
R
2
= coefficient of determination
Table 5. Single predictor of apparent acidogenic value
z
from dietary chemical composition variable
Factor (x) b
0
b
1
R
2
P value
CP (% DM) 12.221 0.119 0.690 <0.0001
Sol. P (% DM) 11.376 0.769 0.163 0.070
Sol. P/CP (%) 5.466 0.157 0.228 0.029
NPN (% DM) 9.596 0.481 0.058 0.291
NPN/Sol P (%) 6.821 0.024 0.019 0.550
Fat (% DM) 9.607 0.229 0.107 0.148
Ash (% DM) 9.269 0.197 0.017 0.575
NDF (% DM) 6.039 0.071 0.141 0.093
ADF (% DM) 5.976 0.187 0.248 0.021
Lignin (% DM) 8.242 0.022 0.0001 0.953
Starch (% DM) 6.930 0.077 0.200 0.042
NFC (% DM) 5.111 0.115 0.425 0.001
Calcium (% DM) 8.624 0.784 0.011 0.652
Phosphorus (% DM) 10.597 4.006 0.231 0.027
z
Apparent acidogenic value (AV) (mg Ca g
1
feed DM) = b
0
+ b
1
(x); where b
0
= intercept, b
1
= slope (increase in AV per unit increase in dietary chemical
composition variables), and x = dietary chemical composition variables.
R
2
= coefficient of determination.
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116 CANADIAN JOURNAL OF ANIMAL SCIENCE
diets that would present a low rumen acid load in dairy cows
and therefore prevent rumen acidosis. There were no differ-
ences in apparent AV and rumen fluid pH change measure-
ments at 24 and 48 h of incubation suggesting that 24 h of
incubation may provide an acceptable measure of feed AV
and rumen fluid pH change. However, further studies are
needed to examine the effect of feed AV on in vivo ruminal
pH changes in dairy cows.
ACKNOWLEDGEMENTS
The authors wish to acknowledge the support of Dairy
Farmers of Ontario, Ontario Ministry of Agriculture, Food
and Rural Affairs and the Natural Sciences and Engineering
Research Council (BWM) for financial support. The techni-
cal assistance of Qian Zhang in the laboratory work is also
gratefully acknowledged.
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z
from dietary chemical composition variables
b
0
b
1
b
2
R
2
P value
Model 1 (all feed classes) 0.81 <0.0001
Intercept 1.628 0.076
NFC (% DM) 0.135 <0.0001
ADF (% DM) 0.236 <0.0001
Model 2 (energy sources) 0.701 0.027
Intercept 0.805 0.772
NFC (% DM) 0.130 0.011
ADF (% DM) 0.356 0.022
Model 3 (fibre sources) 0.844 0.027
Intercept 18.738 0.003
NDF (% DM) 0.146 0.027
Model 4 (protein sources) 0.730 0.014
Intercept 10.686 0.002
CP (% DM) 0.100 0.014
z
Apparent acidogenic value (AV) (mg Ca g
1
feed DM) = b
0
+ b
1
(x
1
) + b
2
(x
2
); where b
0
= intercept, b
1,
b
2
, b
3
= slope (unit increase in AV per unit increase
in dietary chemical composition variable), x
1
and x
2
= dietary chemical composition variables (% DM).
R
2
= coefficient of determination).
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RUSTOMO ET AL. ACIDOGENIC VALUE OF FEEDS. I 117
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