Determination of Rumen Fill, Retention Time and Ruminal

Turnover Rates of Ingesta at Different Stages of Lactation in

Dairy Cows
Gary F. Hartnell and Larry D. Satter

J ANIM SCI 1979, 48:381-392.

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 DETERMINATION OF RUMEN FILL, RETENTION TIME AND RUMINAL
TURNOVER RATES OF INGESTA AT DIFFERENT STAGES
OF LACTATION IN DAIRY COWS 1,2

Gary F. HartneU 3 and Larry D. Satter

University o f Wisconsin, Madison 53 706

SUMMARY within the digestive tract. Grab sampling of

Rumen fill and turnover rates of liquid, feces as compared to total feces collection
grain, and hay in the gastrointestinal tract were resulted in similar estimates of liquid, grain and
measured using rare-earth elements as multiple hay fractions within the digestive tract. Grab
markers in four rumen fistulated cows during sampling of feces as compared to total feces
the dry period and throughout lactation. collection resulted in similar estimates of liquid,
Apparent digestibilities of dry matter, crude grain and hay turnover rates in the reticulo-
protein, neutral-detergent fiber and acid- rumen. Comparisons were made between
detergent fiber were also measured. Hay to stained feed particles and rare-earth elements as
grain ratios of 82.5:17.5, 45:55, 57:43, and markers of ingesta flow.
67:33 (dry matter basis) were fed during the (Key Words: Turnover Rate, Rare-earth Ele-
dry period and during weeks 0 to 12, 13 to 24, ments, Retention Time, Rate of Passage,
and 25 to 44 of lactation, respectively. Apparent Digestibility.)
digestibilities of DM, CP, ADF and NDF were
64.1, 67.2, 68.3, 66.4; 77.2, 70.9, 73.0, 73.7;
41.0, 34.5, 35.6, 38.2;and 51.6, 36.7, 44.0 and INTRODUCTION
47.1 for the four phases of the experiment. Increasing the turnover rate of substrate or
For the dry period, 0 to 12, 13 to 24, and 25 to ingesta has increased yields of rumen microbial
44 weeks of lactation, rumen dry matter and
protein in vitro (lsaacson et al., 1975; Stoutha-
liquid content (kg) were 9.6, 62.1; 16.0, 79.4;
mer and Bettenhaussen, 1973), in sheep (Har-
16.0, 89.3; and 15.7 and 86.0. Mean ruminal
rison et al., 1975; Walker et al., 1975) and in
turnover rates across periods for liquid, grain
steers (Cole et al., 1976). The amount of feed
and hay were 8.1, 4.4, and 3.9% per hour.
protein escaping fermentation might be expected
Experimental period had no effect on turnover
to increase with increased turnover rates of
rate. However, stage of lactation, level of intake
the rumen liquid phase also (Harrison et al.,
and hay:grain ratio all changed with each
1975). In addition, an increased passage rate of
experimental period. The greatest difference in
undigested forage residue out of the rumen may
turnover rate (twofold) was among individual
be associated with increased feed intake with
cows. There were small differences noted high forage rations (Baumgardt, 1970). This,
among stage of lactation periods in total mean
however, is often accompanied by a depression
retention time of liquid, grain and bay fractions in digestibility (Blaxter, 1969).
The suitability of using rare-earth elements
as markers of particulate matter flow in the
gastro-intestinal tract has been discussed
1 Research supported by the College of Agricultural (Hartnell and Satter, 1979). The purpose of this
and Life Sciences, the University of Wisconsin Graduate study was to determine: (1) the range of ingesta
Research Committee, and by Federal Hatch Project
1891. turnover rates and weights of ruminal ingesta
a The technical assistance of Paul Fritschel, Lyle (fill) in dairy cows during the dry period and
Holschbach and Ric Grummer and the assistance in throughout lactation, (2) the effect of stage
neutron activation analysis by Richard J. Cashwell and of lactation on fiber, protein and dry matter
Stephen M. Matusewic are gratefully acknowledged.
3Present address: Allied Mills Inc., Research and digestibilities, (3) the difference between grab
Development Center, P. O. Box 459, Libertyville, IL sampling and total feces collection in deriving
60048. ingesta turnover rates, and (4) the difference
381
JOURNAL OF ANIMAL SCIENCE, Vol. 48, No. 2, 1979

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382 HARTNELL AND SATTER
between stained particle technique and rare-
earth elements as nutrient markers.
M A T E R I A L S A N D METHODS
Four rumen fistulated Holstein cows which
calved within one month of each other were fed
second crop alfalfa hay (IRN-1-00-225) and
grain (table 1) ad libitum in the ratios of
82.5:17.5, 45:55, 57:43 and 67:33 during the
dry period ( - 8 to 0 weeks), early lactation (0
to 12 weeks), midqactation (13 to 24 weeks),
and late lactation (25 to 44 weeks), respectively.
The cows were first fed the 82.5:17.5 hay:grain
ratio 8 weeks prior to calving. Enough alfalfa
hay was harvested from one field to last the
entire experiment. Hay and grain were analyzed
for crude protein using the Kjeldahl procedure
and for acid- and neutral-detergent fiber using
the procedure of Goering and Van Soest, 1970.
One-third of the daily grain allotment was
fed at 0400 hr before the morning milking,
again at mid-morning (1030 hr) and again at
1700 hr after the afternoon milking. Feed and
water intakes were recorded daily. Water
intakes were measured using water meters
placed in the water line between the drinking
cup and the main water line. Body weights were
taken once a week at 0745 hours. Weight of
total rumen contents was measured four times
over a 4- to 6-week period in the middle of each
phase. Two hours after the mid-morning
feeding, the reticulorumens were manually
emptied. The rumen contents were weighed,
and numerous samples were taken as the
contents were placed back into the rumen.
Digesta samples were composited and dried in a
forced air oven at 60 C for 72 hr to determine
dry matter content. Digestibility. and rate of
passage measurements were performed during a
2.5-week sampling period in the middle of each
phase as illustrated in figure 1.
Samarium (Sm) and lanthanum (La) solutions
were prepared b y dissolving 1.16 g of samarium
oxide (99.9% pure) or 1.17 g of lanthanum
oxide (99.9% pure) 4 in 4 ml of concentrated
hydrochloric acid and diluting with water to 30
milliliters. The cerium solution was prepared by
dissolving 3.92 g of ceric ammonium nitrate
4Research Chemicals, P. O. Box 14588, Phoenix,
AZ 85603.
s G. Frederick Smith Chemical Company, Colum-
bus, OH.
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TABLE 1. COMPOSITION OF THE
GRAIN MIXTURE
Ingredient I RN a Percent
Ground shelled corn 4-02-931 50.0
Ground oats 4-03-309 21.7
Soybean oil meal (44%) b 5-04-612 15.0
Beet pulp 4420-669 10.0
Dried cane molasses 4-04-695 1.2
Dicalcium phosphate 6-01-080 1.0
Trace mineralized saltC ... 1.0
Vitamin ADE premixd . . . . 1
aAtlas of nutritional data on United States and
Canadian feeds. 1971. National Academy of Sciences,
Washington, DC.
bLinseed meal (5-02-048) substituted for soybean
meal during the lactation phases of the experiment.
CThe mineral mix contained: NaCI (95-99%); Mn
(>.20%); Ferrous (Fe), (>.16%); Ferric (Fe), (>.14%);
Cu (>.033%); Zn (>.010%); I (>.007%); and Co
(>.005%).
dvitamin premix contained 2,000,000 IU of
vitamins A and D and 200 IU of vitamin E per
kilogram.
(99.9% pure) s in 10 ml of water and diluting
with water to 30 milliliters. The resulting
solutions contained approximately 1 g of
element per 30 milliliters. Solutions were stored
in polyethylene containers until needed.
The marked hay was prepared by mixing .5
kg of crystal violet stained alfalfa hay (Balch,
1950) with .5 kg of unstained alfalfa hay. The
mixture was sprayed with 30 ml (approximately
1 g of element) of each of the rare-earth solu-
tions with each cow receiving a different
combination of elements. The reason for using
different combinations of elements was to
verify that all o f the elements behaved similarly.
Stained hay was included to allow comparison
o f the two methods of marking. After spraying,
the hay was air dried and placed into plastic
garbage bags until needed for feeding. Marked
grain was prepared as described for the hay
except .25 kg of basic fuchsin stained grain
(Balch, 1950) was mixed with .75 kg of un-
stained grain before spraying with La, Ce
and/or Sm solutions. Thus, the marked meal
fed each cow consisted of 1 kg of treated hay
and 1 kg of treated grain. One liter of Cr-EDTA
(2.6 to 3.0 g Cr) prepared according to the
procedure of Binnerts et al. (1968) was placed
into the rumen via the fistula at the time of
feeding the marked meal. Marked hay and
 RUMEN FILL, RETENTION AND TURNOVER ON LACTATION 383

Day
i 2 3 4 5 6 7 8 9 i0 Ii 12 13 14 15 16 17

A 71
B B

c ;hi D C "~1

r - ~1

Figure 1. Description of 2.5 week sampling period during which time all ingesta and feces samples were
collected. A) Feed and water intakes were recorded daily. B) Marked hay and grain were fed with 1 liter of
Cr-EDTA at 1030 hours. C) Grab samples of rumen contents were taken at O, 6, 12, 18, 24, 30, 36, 48, 60, 72,
96, 120 and144 hr after feeding the marked meal. Grab samples of feces were taken at O, 12, 18, 24, 30, 36, 48,
60, 72, 84, 96, 108, 120, 144 and 168 hr after feeding the marked meal. D) Measured rumen fill. Three addition-
al measurements of rumen fill were obtained just prior to or following this 2.5-week period. E) Attached feces
collecting apparatus. F) Attached feces bag. The bags were changed every 6 hr and the 12 hr collections com-
posited through 7 days. G) Removed feces collecting apparatus.

grain that was not consumed by the cow after
45 min were manually broken into 10 cm
pieces and placed directly into the rumen via
the fistula. Usually this amount did not exceed
10 to 20% o f the total marked feed.
Grab samples of rumen contents, consisting
of a composite of samples taken from six
different locations in the rumen, and grab
samples of feces were weighed, dried at 60 C
for 48 hr, weighed, and ground through a 40
mesh screen using a Wiley mill. Portions of the
ground samples (2.5 to 4.0 g) were weighed
into 4-dram polyethylene vials for neutron
activation analysis of Cr, Sm, Ce and La. In
obtaining the feces, the animal was induced to
defecate and the last portion of feces excreted
was taken as the sample.
Total feces collections were performed using
the feces collecting apparatus described by
Gorski e t al. (1957). Using this apparatus, the
cows could go outside for exercise and be
milked in the parlor during the collection. No
urine was collected. Twelve-hour feces compos-
ites and grab samples of feces for the 7-day
collection period were prepared for neutron
activation analysis of Cr, Sm, Ce and La as
described b y Hartnell and Satter (1978). The
ground dried feces from the total feces collec-
tion were composited into 24-hr samples. These
samples were analyzed for crude protein
(Kjeldahl method) and acid- and neutral-_
detergent fiber (Goering and Van Soest, 1970).
The stained particle technique employed was
similar to that used by Balch (1950) and Castle
(1956) except that .1 g of dried ground feces
was spread out over a 12.5 cm grid and then
counted with the aid of a microscope. Wet feces
were not washed over a piece of cheesecloth
and counted, as Balch recommended, since
cheesecloth withholds only those particles
of diameter greater than the pores o f the cloth,
whereas no particles were lost using a dried
ground sample.
Liquid, hay and grain ruminal turnover rates,
total mean retention times and transit times
were calculated by fitting a two compartmental
model (Brandt and Thacker, 1958) to the fecal
excretion data (grab samples and total feces
collection) using the procedure o f Grovum and
Williams (1973b).
A completely randomized design was used in
calculating the analysis o f variance. The means
were compared using Duncan's new multiple
range test (Duncan, 1955).
RESULTS A N D DISCUSSION
Dry matter, crude protein, neutral-detergent
fiber and acid-detergent fiber contents in hay
and grain which were fed to the cows are
reported in table 2. Table 3 contains b o d y
weight, milk production, water and feed intake

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384 HARTNELL AND SATTER

TABLE 2. DRY MATTER (DM), CRUDE PROTEIN
(CP), NEUTRAL-DETERGENT FIBER (NDF), AND
ACID-DETERGENT FIBER (ADF) ANALYSIS
OF RATION INGREDIENTS
Feedstuff DM CP NDFa
(%)
Hay 89.9 17.2 49.4
Grain 90.1 18.3 17.5
aNeutral-detergent fiber (Goering and Van Soest,
1970).
bAcid-detergent fiber (Goering and Van Soest,
1970).
and ration digestibility data for each phase of
the experiment. In evaluating the results, it
must be kept in mind that the effects of feed
intake, hay to grain ratio and stage of lactation
are confounded.
Water and dry matter intakes were lower
(P<.05) (25% and 43%) during the dry period
but were different (P<.05) during the periods
of milk production. It was observed that during
very hot (T>32 C) days cows would reduce
their dry matter consumption without changing
water intake.
Apparent dry matter digestibility found in
the dry period was less (P<.05) than the three
periods in which the animals were lactating.
This was probably due to the lower proportion
of grain fed during the dry period. Apparent
digestibility of crude protein increased (P<.05)
from 70.9% after parturition to 77.2% during
the dry period. Apparent digestibility of
neutral-detergent fiber increased (P<.05) as the
proportion of the hay in the diet increased.
ADFb This was also evident for the acid-detergent
fiber. Campling (1966) reported that addition
of grain to the ration increased dry matter
34.5 digestibility, but decreased crude fiber digesti-
8.3
bility.
Measurements of rumen fill are presented in
table 4. There was 40% less dry matter in the
reticulo-rumen during the dry period compared
to the milking periods. The liquid portion of
the rumen contents varied from 62.1 kg during
the dry period to 89.3 kg during weeks 13 to
24 of lactation. There was a 10 kg increase in
total ruminal ingesta weight from the middle of
the 0 to 12 week period to the middle of the 13
to 24 week period. On average, total rumen fill
was closely related to dry matter intake. It is
important to bear in mind the changes in
ingesta weight or gut fill when examining
changes in overall body weight during early
lactation. A cow may actually be losing body
tissue during this time, but the increase in gut
fill (24 kg in this study) would obscure this.
Therefore, body weights must be used carefully
when trying to assess depletion of body stores
during early lactation.
When the rumens were emptied, it was noted
that among the four cows, 2273 seemed to have
the smallest rumen, and 2136 the most spacious
rumen. The dry matter content in each was

TABLE 3. BODY WEIGHT, MILK PRODUCTION, WATER AND DRY MATTER INTAKE, AND
APPARENT DIGESTIBILITY OF RATION COMPONENTS AT DIFFERENT
STAGES OF LACTATION

Stage of lactation (weeks)

(Dry)
Component -8-0 0-12 13-24 25-44 SE

Forage:Grain 82.5:17.5 45:55 57:43 67:33 ...

Body weight (kg) 700 606 628 656 ...
Milk production (kg/day) 0 28.5 24.4 16.2 ...
Water intake (kg/day) 60.5 a 82.9b 83.8b 78.6b 2.7
Dry matter intake (kg/day) 10.8a 16.9b 19.8b 17.6b .9
Apparent digestibility, %
Dry matter 64. la 67.2b 68.3 b 66.4 b .6
Crude protein 77.2c 70.9 a 73.0ab 73.7 b .7
Acid-detergent fiber 41.0 34.5 35.6 38.2 1.4
Neutral-detergent fiber 51.6b 36.7a 44.0b 47.1 b 3.1

a'b~CMeansin the same row which do not have a common letter in the superscript are different (P<.05).

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 RUMEN FILL, RETENTION AND TURNOVER ON LACTATION 385

TABLE 4. RUMEN FILL AT D I F F E R E N T STAGES OF LACTATION

Stages of lactation (weeks)

(Dry)
Component -8-0 0-12 13-24 25-44 SE

(kga)
Dry matter 9.6 b 16.0c 16.0 c 15.7 c .9
Liquid 62.1 b 79.4 c 89.3 d 86.0 ~d 3.2
Total 71.7 b 95.4c I05.3 c 101.7 c 3.1

aEach value is a mean of four cows.

b'C'dMeans in the same row which do not have a c o m m o n letter in the superscript are different (P<.05).

approximately the same, even though cow 2273 differences among cows, particularly in liquid
had a very congested, filled reticulo-rumen and fill of the rumen and in turnover of liquid
cow 2136 had ample space above the ingesta ingesta (table 5). Cow 2136 had up to 30 kg
for additional material. There were large more liquid than cow 2273 during the lactation

TABLE 5. INDIVIDUAL COW DATA FOR RATE OF PASSAGE MEASUREMENTS DURING PERIODS
OF LOWEST AND HIGHEST DRY MATTER INTAKES

Stage of lactation (weeks)

(Dry) -8-0 (Lactating) 13-24
Cow identification
Parameter 2273 2149 2136 2146 2273 2149 2136 2146

Body weight, kg 697 710 648 747 644 620 632 615
Milk production, kg/day 999 . . . . 23.3 21.6 24.3 28.2
Dry matter intake kg/day 11.2 " 10.4 10.0 11.6 21.8 17.8 19.2 20.3
Water intake, kg/day 53.1 73.6 59.5 55.8 86.3 77.7 85.7 85.6
Rumen fill, kg
Dry matter 7.6 9.7 10.9 10.3 16.4 14.5 17.8 15.1
Liquid 50.7 70.5 70.5 53.5 80.7 83.0 113.8 79.8
K1, %/hra
Liquid 8.9 4.7 8.0 9.3 12.0 7.2 6.0 10.1
Grain 4.8 3.9 3.1 4.1 5.5 4.3 3.6 5.0
Hay 3.9 3.3 2.9 3.4 4.5 3.6 2.5 3.2
K 2 , %/hrb
Liquid 30.8 109.7 12.5 33.0 52.5 84.0 76.4 47.4
Grain 11.3 11.9 6.5 14.1 27.4 21.7 40.0 26.7
Hay 7.5 8.3 5.1 9.7 17.4 9.1 19.0 12.1
TMRT, hr c
Liquid 26.0 33.7 31.9 22.5 21.5 26.6 34.3 23.1
Grain 41.5 45.5 59.1 42.6 33.7 39.7 48.0 35.3
Hay 50.7 53.7 65.9 50.9 39.4 50.6 62.7 50.1
TT, hr d
Liquid 11.5 11.2 11.5 8.8 11.2 11.5 16.2 11.0
Grain 11.8 11.8 11.8 11.3 11.8 11.8 17.5 11.5
Hay 11.8 11.8 11.8 10.8 11.5 11.5 17.0 10.8

ak 1 - ruminal turnover rate.

bk 2 - thought to be turnover rate of contents in the hindgut or cecum and proximal colon or
artifact due to mixing.
CTM RT - total mean retention time.
dTT - time for first appearance o f marker in feces.

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386 HARTNELL AND SATTER
periods. Water intake was about the same for
these two cows during lactation. Differences
between cows in rumen liquid volume may be
due to differences in saliva production, liquid
turnover rate and/or ruminal capacity. If faster
fluid turnover rates in the rumen are correlated
with greater efficiency of microbial growth
(Isaacson et al., 1975; Walker et al., 1975), then
cows 2273 and 2146 might be expected to
have a larger supply of microbial protein than
cows 2149 and 2136.
Dry matter intake as well as dry matter
content in the rumen were reduced during the
dry period, and the question might be raised
whether the increase in size of the fetus reduced
rumen fill and limited feed intake. In this
study, rumens were emptied at 8, 5 and 2 or 1
week prior to parturition. There were no
differences in the amount of dry matter or
liquid in the reticulo-rumens at any time during
the dry period. Lamberth (1969) reported
nonsignificant differences in rumen fill between
nonpregnant and pregnant twin heifers.
Samarium (Sm), cerium (Ce)and lanthanum
(La) were used as particulate markers because
they adsorb tenaciously to feed particles, are
inexpensive, have nonoverlapping decay patterns
upon activation, and have sufficiently long half
lives to enable counting 7 to 10 days after
neutron activation (Hartnell, 1977; Hartnell
and Satter, 1979). Therefore, excretion of
the markers should be representative of the diet
component marked. The exception might be
that fraction of marker which was adsorbed
onto particles which were subsequently digested
away. In general, close to 100% of the elements
were recovered in the feces, with the extreme
range of recovery equaling 92% and 115%.
Urine and milk were monitored for the eleme-
nts throughout the experiment, and no rare-
earth element was detected.
In calculating and measuring ingesta turnover
rates and total mean retention times in the
gastrointestinal tract, the method of Grovum
and Williams (1973b) was used to fit the two
compartmental model described by the follow-
ing equation (Brandt and Thacker, 1958) to the
fecal excretion data:
y = Ae-ka (T-TT) _Ae-k2 (T-TT) T>TT
y =o T-<<TT
where y is the concentration of marker in the
feces; kl represents the turnover rate of marker
from the reticulo-rumen; T is the time elapsed
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since feeding of the marked meal; TT denotes
the time of first appearance of marker in the
feces; A is biologically undefined; and k2 is
thought to be related to the turnover rate of
marker post ruminally or in the cecum and
proximal colon (Grovum and Williams, 1973b),
or to be an artifact which is dependent upon
mixing within the rumen (Leverett et al.,
1977). The importance of this approach is that
ruminal turnover rates of ingesta can be calcu-
lated based on fecal excretion of marker.
Computer programs (Hartnell, 1977) were
used in calculating the rate constants and
transit times based on the procedure used by
Grovum and Williams (1973b). Chi-square was
used as an index of dispersion in determining
the equation parameters that best fit the
semiqog plot of actual excretion data.
The turnover rates and total mean retention
times for liquid, grain and hay at various stages
of lactation were obtained by the following
methods of calculation: 1) determining the best
fit of the two compartmental model to the data
obtained by grab sampling feces; 2) determining
the best fit of the two compartmental model to
the actual fecal excretion curve using the total
feces collection data; 3) determining the best fit
of the two compartmental model to the cumu-
lative percentage recovery curve using the total
feces collection data; 4) determining the mean
retention time using the stained particle tech-
nique; and 5) grab sampling of rumen contents
to determine the rate of marker disappearance
(k~) from the rumen.
Method 1 is perhaps the preferred method of
calculation, and it is illustrated with the follow-
ing example. Excretion of the hay component
in feces using La as the marker for cow 2146,
phase 4, is presented in table 6. First, the
concentration of La detected in the feces was
corrected by substracting background levels.
Secondly, the concentration values were then
transformed into their respective natural
logarithms (table 6). If the rate of removal of
marker from the reticulo-rumen is exponential
with time, then the plot of the natural logar-
ithm of the marker concentration versus time
should be linear after a certain amount of time,
normally 24 to 36 hr in this experiment.
y=At e "kt T
l n y = I n A ~ -k~ T
The absolute value of the regression coeffi-
cient (slope) obtained from the natural logar-
 RUMEN FILL, RETENTION AND TURNOVER ON LACTATION 387

TABLE 6. CONCENTRATION OF LA IN GRAB TABLE 7. DETERMINATION OF K1 VALUES

SAMPLES OF FECES COLLECTED IN PHASE 4 USING VARIOUS SETS OF SAMPLES IN A
FOR COW 2146 AFTER FEEDING A MEAL LINEAR REGRESSION ANALYSISa
CONTAINING LA MARKED HAY (GRAB SAMPLINGMETHOD)

Sampling La La Set of sample

Sample time feces feces values usedb r2 lnAl A1(ppm) k1
No. (hr) (ppm) (ln ppm)
5-14 .986 5.539 254.4 .03469
1 0 1.63 6-14 .986 5.634 279.8 .03559
2 12 .56a 2.607 6-13 .982 5.520 249.5 .03388
3 18 48.67 3.885
4 24 65.69 4.185
5 30 76.40 4.336 alny = lnA, - k , T; where y is the concentration of
6 36 59.09 4.079 marker in the excreted feces, T is the sampling time,
7 48 56.71 4.038 A, is the antilogarithm of the intercept of the regres-
8 60 35.87 3.580 sion line on the Y axis and k 1 is the absolute value of
9 72 22.20 3.100 the regression coefficient (slope): r 2 is the coefficient
10 84 14.86 2.699 of determination.
11 96 9.29 2.229 bsample values used are from table 6.
12 108 7.40 2.001
13 120 3.64 1.292
14 144 1.42 .351
constant. TT is initially set to a time which is
asamples 2-14 have had the zero time concentra- estimated to be before the time of first appear-
tion subtracted from them. The zero time concentra-
tion was considered background. ance of marker. In table 6, La was detected at
12 hr, therefore an initial TT of 8.25 hr might
be selected. Thus the actual T r should be
between 8.25 hr and 12.00 hr. While holding kl
ithms of the marker concentration in the feces and TT constant, k2 is varied. The sampling
in the linear portion of the curve is k~. The y times (12, 18, 24, 30, 36, 48, 60, 72, 84, 96,
intercept of the regression line yields the value 108, 120, 144 and 150 hr) are then computed
for l n A , . The data point with which to start and stored. The A value is computed as being
the linear regression analysis was selected so equal to A l e - k 1 T T (Grovum a n d Williams,
that the values in the rising or peak portion of 1973b). Then at each sampling time through
the curve did not appear to bias the regression the final sampling time read in, the natural
coefficient. Generally, the highest or peak value logarithm of fecal marker concentration will be
was used as the starting point (sample 5 in table calculated using the following equation:
6). Next, various sets of sample values were
used in the linear regression analysis to obtain In y = ln(Ae"kl (T-TT)--Ae-k2 ( T - T T ) ) T > T T
values for kl and A1 (table 7). Initially, all of
the values from the peak onward (samples 5 to In y = 0 T~ TT
14) were used, then values near the peak and at
the tail end of the curve were deleted and the The predicted In y for each sampling time is
regression parameters recalculated. The resuhing compared with the In (marker concentration)
kl and A1 values are presented in table 7. The actually detected in the feces by using chi-
coefficient of determination for each regression square. The k I , k2, TT and chi-square are then
line was very high (.986). stored. Still holding kl and TT constant, k2 is
A computer program (HartneU, 1977) was incremented by .01 units and a new set of In y
used in determining the best k2, TT and A for values are calculated. If the chi-square value is
each kl value. The k l , AI, an initial k2 value less than the chi-square value calculated with
(generally kx + .03) was used) and the final the previous k2, then the current k l , k2, TT
excretion time were read into the computer and chi-square values are stored and k2 is again
along with the natural logarithms of the marker incremented by .01 until k2 has been incre-
concentrations in the feces (table 6, column 4). mented 52 times. If the chi-square value is
As an example, .03469(kl ), 254.4(A1), .06460 greater than the stored value, the parameters
(k2) and 144 hr (tables 6 and 7) would be read are not stored and the program proceeds to
into the computer. The program holds kl increment k2 and calculate new values. Afte~

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388 HARTNELL AND SATTER

k2 has been increased 52 times as indicated in
the do loop, the stored k l , k2, TT, chi-square
and predicted in y at each sampling time are
printed out.
The computer then increments TT by .25 hr
while holding kl constant. The program starts
over again with the same kl and the new TT
value held constant and the k2 varied. This
procedure is repeated until a predetermined TT
value is reached. This predetermined time is the
sampling time in which marker is first detected
in the feces. Thus, the computer printout
contains a list o f parameters with k 1 being
constant and the best k2 determined with each
increment of TT (table 8). If TT was varied
from 8.25 to 1200 hr at .25-hr increments,
there would be 16 parameters printed out
(table 8). Of the parameters printed out, the set
with the minimum chi-square value would be
the one that resulted in the best fit of the
model to the data for that particular k l . In the
example, this would be .03469(kl ),
.01469(k2), 10.75 hr (TT) and .133 (chi square)
as presented in table 8. When TT was equal to
1200 hr, the chi-square was lower than when
TT was equal to 10.75 hr (table 8). This oc-
curred because when TT equalled 12.00 hr, the
predicted value was set to zero and thus no
comparison was made between the predicted
and actual values. If the actual value would
have been zero, then the chi-square would have
been correct.
So far, k2 and TT have been varied with kl
being held constant. In comparing the parame-
ters obtained when the three kl values from
table 7 were used, .03559 was the best k I value
to be used. Table 10 compares the predicted
values using the three sets of parameters in
Table 9 versus the i n (actual excretion of
marker) detected. Therefore, .03559(kl),
.09560(k2), 10.75 hr (TT) and 190.8 ppm (A)
were the parameters that resulted in the overall
best fit. These parameters were then used
in calculating turnover time (1/kl ; 1/k2), total
mean retention time (TT + 1/kl + 1/k2) and
half-rimes (.693/kl ;.693/k2).
The procedure used for methods 2 and 3 was
essentially the same as that used for method 1
(Hartnell, 1977), except in method 3 the
cumulative percentage of marker recovered was
predicted according to the equation of Grovum
and Phillips (1973):

PC = 100[(k2 - kl + k l e - k 2 ( T - T T )
--k2e - k l (T--TT))/(k2 -- kl )] when T > T T
TABLE 8. VALUES OF K 2 AND TT RESULTING
FROM THE BEST FIT OF THE TWO COMPART-
MENTAL MODEL TO THE DATA WHEN TT PC=0 whenT<TT
WAS INCREMENTED AND K 1 HELD
CONSTANT (GRAB SAMPLING METHOD) where PC is the cumulative percentage of
excreted marker and the other symbols are the
Set kt k2 TT Chi-square same as described earlier.
Figure 2 demonstrates how well this model
1 .03469 .08469 8.25 .351 actually fits the total feces collection data for
2 .03469 .08469 8.50 .308 the cumulative excretion of Cr (liquid), Sm
3 .03469 .08469 8.75 .285 (hay) and La (grain). The symbols represent
4 .03469 .08469 9.00 .263
5 .03469 .08469 9.25 .241 actual data and the smooth curves represent the
6 .03469 .08469 9.50 .221 best fit of the model to the data. The cumulative
7 .03469 .08469 9.75 .203
8 .03469 .09469 10.00 .181
9 .03469 .09469 10.25 .159
10 .03469 .09469 10.50 .146 TABLE 9. COMPUTER CALCULATED BEST FIT
11 .03469 .10469 10.75 .133 a VALUES FOR K2, TT AND A AT THREE
12 .03469 .11469 11.00 .134 DIFFERENT K 1 VALUES (GRAB
13 .03469 .13469 11.25 .157 SAMPLING METHOD)
14 .03469 .16469 11.50 .218
15 .03469 .31469 11.75 .341
16 .03469 .11469 12.00 .126 b Set k 1 ka TT A Chi-square

aMinimum chi-square. (hr) (ppm)

bWhen TT equals 12, there is no comparison made 1 .03469 .10469 10.75 175.2 .133
between the actual value and predicted value because 2 .03559 .09560 10.75 190.8 .106
the predicted value is set to equal zero. Thus the 3 .03388 .10388 10.75 173.3 .186
chi-square is incorrect if the actual value is not zero.

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 RUMEN FILL, RETENTION AND TURNOVER ON LACTATION 389

TABLE 10. COMPARISON OF THE COMPUTER the large. Thus the excretion curve of Sm is
PREDICTED VALUES USING THREE reflective of the mean rate of passage for the
DIFFERENT K1 VALUES WITH THE
ACTUAL VALUES OF LA hay component.
DETECTED IN THE FECES Calculations used in method 4 for obtaining
(GRAB SAMPLING METHOD) the R values are described by C a s t e (1956).
Method 5 simply assumed that feed components
Calculated La excreted passed out of the reticulorumen at an exponen-
Actual, La
. in feces usingk. 1' k 2 ' A tial rate (Y = A e - k t ) , and that a linear regression
Sample Sampling excreted and TT values m table 9. of a plot of l n Y versus time would give k, the
no. time in feces 1 2 3 rate of disappearance from the rumen. Examples
of calculations for each of the five methods are
In (ppm) available (Hartnell, 1977).
1 o The turnover rates and total mean retention
2 12 2[607 2[643 2[579 2[634 times (TMRT) for the liquid, grain and hay
3 18 3.885 3.993 3.951 3.989
4 24 4.185 4.203 4.179 4.203 during the dry period and various stages of
5 30 4.336 4.197 4.188 4.203 lactation are presented in table 11. The values
6 36 4.079 4.103 4.104 4.113 are means obtained by averaging results from
7 48 4.038 3.797 3.812 3.817 cows obtained by method 1 (grab sampling of
8 60 3.580 3 . 4 2 5 3.445 3,455
9 72 3.100 3.027 3 . 0 4 5 3.067 feces). This would perhaps be the method of
10 84 2.694 2.616 2 . 6 3 1 2.668 choice in most situations.
11 96 2.229 2.206 2.210 2.265 If we assume that kl obtained from the two
12 108 2.001 1 . 7 9 1 1.786 1.860 compartmental model represents the turnover
13 120 1.292 1.276 1 . 2 6 1 1.454
rate of the marked fraction within the reticulo-
14 144 .351 .543 .507 .641
rumen (Grovum and Williams, 1973b; Grovum
and Phillips, 1973), then turnover rates of the
liquid, hay and grain fractions did not signifi-
cantly differ with stage of lactation. Stage of
percent recovery curves (figure 2) might be lactation, of course, is confounded here with
considered to represent the average of a family level of feed intake and forage to concentrate
of curves for each of the hay, grain and liquid ratio. High levels of dry matter intake might be
fractions. Since the hay particles that were expected to increase turnover rate. The possible
sprayed with Sm were of different particle reduction in cellulose hydrolysis with high grain
sizes, one would expect a different rate of feeding would tend to decrease the turnover
passage for the small particles as compared to rate. Thus, these factors may balance each
other out. Another factor that may be involved
is calorie demand. Kennedy e t al. (1976)
IO0
reported that sheep challenged with an increased
calorie demand as a result o f cold exposure had
an increased rate of digesta passage. It is con-

t
ceivable that in early lactation, when'the cow is
Lu
mobilizing body tissue to supply calories, that a
similar situation might exist in the dairy cow.
Either stage of lactation did not have a marked
effect on turnover rate, or the high grain level
~4Q 9 LIQUID
and its adverse effect on cellulose hydrolysis
2 tended to compensate for the influence that
higher levels of dry matter intake and calorie
demand would be expected to have on increasing
turnover rate. Whatever the explanation,
% 2 40I I 60I ~80~-1 IooI I
t20
I 140
I I ,/~
ruminal turnover rates of ration components
TIME AFTER FEEDING MARKED MEAL (HR)
were relatively unchanged over the period of
Figure 2. Cumulative percentage recovery curves of time the cows were lactating.
Cr (liquid), Sm (Hay) and La (Grain) where the
symbols (0, +, zx) represent the actual data and the The k2 values up to this point in time have
smooth curves represent the best fit of the model to uncertain biological meaning. Originally, T~Ak2
the actual data. was thought to be the half-time of marker in

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390 HARTNELL AND SATTER

TABLE 11. COMPONENT TURNOVER RATES, TOTAL MEAN RETENTION TIMES AND TRANSIT
TIMES FOR RATION CONSTITUENTS AT DIFFERENT STAGES OF LACTATIONa

Stage of lactation (weeks)

(Dry)
Component -8-0 0-12 13-24 25-44 SE

Hay:Grain 82.5:17.5 f 45:55g 57:43 h 67:33 i ...

k~, %/hrb
Liquid 7.7 9.6 8.8 8.7 1,54
Grain 4.0 5.2 4.6 4.1 .51
Hay 3.4 3.8 3.4 3.5 ,39
k2, %/hr c
Liquid 46.5 45.5 65.1 35.4 11.57
Grain 11.0f 31.1 h 28.9gh 15.8fg 4.70
Hay 7.7f 13.1g 14.4g 8.8 f 1.47
TMRT, hrd
Liquid 28.5 27.7 26.4 28.6 3.68
Grain 47.2 37.9 39.2 45.1 4.34
Hay 55.3 49.9 50.7 54.4 4.90
"IT, hre
Liquid 10.8 12.6 12.5 12.4 1.2
Grain 11.6 13.1 13.1 12.7 1.3
Hay 11.5 13.9 12.7 13.9 1.4

acalculated by method 1.
bk I - ruminal turnover rate.
Ck2 - thought to be turnover rate of contents in the hindgut or cecum and proximal colon or artifact
due to mixing.
dTMRT - total mean retention time.
eTT - time of first appearance of marker in the feces.
f'g'h'iMeans in the same row which do not have a common letter in the superscript are different (P<.05).

the hind gut, but when Grovum and Williams
(1973b) injected markers into the rumen and
collected feces they found that T%k: on the
average was about 66% of the half-time (Ty2a)
computed by injecting the marker into the
abomasum. One would expect the two methods
to give similar values. If there was a diurnal
pattern of digesta mixing in the caecum and
proximal colon, Ty2k~ might be affected
differently from TY2a values because TtAk 2
was determined over a shorter period of time.
When Ty~k2 was measured in a model digestive
tract, there was no difference between it and
the expected value (Grovum and Phillips,
1973) indicating that something different was
happening in vivo. The effect of time of injec-
tion has not been evaluated. However, Grovum
and Williams (1973b) found good fits between
the concentrations of ~44Pr and S lCr-EDTA
in feces and the calculated curves which indi-
cates that T%k 2 does describe the way marker
and digesta passed through the hind gut. Since
the half times of 144 Pr and s 1Cr-EDTA were
relatively small in the abomasum, 37 and 17
min, compared with those in the caecum and
proximal colon, 413 and 406 min (Grovum and
Williams, 1973a), there is reason to associate
T~k2 with the kinetics of digesta flow through
the caecum and proximal colon. The changes in
Ty2k2 may therefore reflect differences in the
times that digesta spends in the caecum and
colon, but the values for average retention
times (1/k~) may n o t be the true times that
digesta actually spends in these organs (Grovum
and Williams, 1973b). Finally, the differences
of magnitude between Ty2k2 and T89 in vivo
may be due to imperfect mixing of digesta in
the caecum and proximal colon of the sheep
(Grovum and Williams, 1973a).
Leverett, Marls and Ellis (1977) suggested
that k2 is an artifact due to mixing in the
reticulo-rumen, or in other words, that mixing
in the rumen represents the second compart-
ment (k2) with rate of passage out of the
rumen as the first compartment (ki). However,
there is perhaps more evidence at this time to

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 RUMEN FILL, RETENTION AND TURNOVER ON LACTATION 391

suggest that k2 reflects retention of ingesta TABLE 12. COMPARISON OF METHODS FOR
in a post ruminal compartment than of a DETERMINING RATE OF PASSAGE
PARAMETERS
mixing phenomenon.
In this experiment, k2 values tended to be
Methoda
greatest for hay and grain when 55% of the
ration was grain, and decreased as the propor- Parameters 1 2 3 4 5
tion of grain in the ration was reduced. The ka, %/hr
liquid turnover rate remained fairly constant Liquid 8.7 c 7.8 c 8.0c 9.7 d
except for an unexplainable large value during Grain 4.5 4.4 4.4 4.4
the third phase of the experiment. The k2 value Hay 3.5c 3.7 cd 3.7cd 3.9 d
k2, %/hr
was always found to be greater than kl which Liquid 48.1 d 28.1 c 46.3 d
agrees with Grovum and Williams (1973b). The Grain 21.7 24.3 31.2
k2 value has limited meaning in this experiment Hay 11.0 13.7 12.8
because it is based on only two to three samples TMRT, hr R-valueb
or data points. To obtain more reliable estimates Liquid 27.8 e 25.1 d 22.1c
Grain 42.3d 40.5 d 36.0c 41.5 d
of k2, at least six samples should be obtained Hay 52.6d 48.6 d 45.3 c 53.6d
during the first 24 hr after feeding the marked
meal. aMethod 1 : determining the best fit of the model
The total mean retention time (TMRT) was to the data obtained by grab sampling feces. Method
calculated as TMRT = TT + l/k1 + l / k 2 . The 2: determining the best fit of the model to the actual
R-value was found to be at least 97.8% of the marker excretion curve (total feces collection).
Method 3: determining the best fit of the model to
TMRT value. The TMRT of liquid and hay the cumulative percent recovery curve (total feces
fractions did not differ significantly from collection). Method 4: stained particle technique.
period to period (table 11). Grain tended to Method 5: grab sampling of tureen contents.
spend more time in the digestive tract when the bcalculated as described by Castle (1956).
ration contained relatively small amounts of c'd'eMeans in the same row which do not have a
grain. common letter in the superscript are different (P<.05).
The time of first appearance (TT) of liquid,
grain or hay in the feces were n o t different
(P<.05) with regard to stage of lactation,
however they tended to be shorter during the rare-earth elements in either feces grab samples
dry period. or the two total feces collection methods.
Table 12 contains comparisons of turnover There was no difference among the grab sam-
rates obtained by using feces grab sampling pling method, stained particle method and the
(method 1) or the two methods (methods 2 and total feces collection method using the best fit
3) using the total feces collection data. The of the I n (marker concentration) to the actual
values are means of the data collected over all value except for the liquid component. The
periods of the experiments. The kl values total feces collection method using the best fit
calculated from these data are also compared of cumulative percentage recovery values to the
with the k value obtained from the rumen grab actual cumulative percentage recovery values
sampling method (method 5). There were no (method 3 in table 12) tended to yield lower
differences (P<.05) between the fecal grab total mean retention time. The reason for the
sampling method and the total feces collection difference is yet unexplained.
methods in determining kl or k2. The signifi- On the basis of the relative differences
cantly lower k2 value for the liquid phase using among the methods of analysis, grab sampling
method 2 is unexplainable. Rumen grab sam- of feces from the rectum would be the desired
piing tended to yield slightly higher values for method to use. The important point with this
k l , especially for the liquid component. The method is that the animal should be induced to
reliability of the rumen grab sample values is defecate and the last excrement taken as the
limited because representative samples were sample to avoid any bias due to storage of feces
difficult to obtain and a limited number of in the rectum. Using this method with rare-
samples (3 or 4) were taken during the descent earth elements as multiple component markers
of the exponential curve. within a ration, one can use more animals
The TMRT values were used in comparing without the laborious task of making a total
the stained particle technique with the use of feces collection. The method also enables one

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392 HARTNELL AND SATTER
t o calculate t h e r u m i n a l t u r n o v e r rates for each
c o m p o n e n t m a r k e d a n d t h e overall m e a n
retention time just from the marker concentra-
t i o n s in t h e feces. It is suggested t h a t at least 6
t o 8 feces samples b e c o l l e c t e d w i t h t h e first 24
h r if a reliable e s t i m a t e o f k2 is t o b e o b t a i n e d .
T h e k2 value m u s t b e viewed w i t h c a u t i o n ,
h o w e v e r , u n t i l its biological m e a n i n g is b e t t e r
described.
T h e rare-earth e l e m e n t s are easily a p p l i e d t o
t h e r a t i o n w i t h o u t a l t e r i n g t h e c h e m i c a l or
physical p r o p e r t i e s o f t h e r a t i o n , w h i c h o f
course h a p p e n s w h e n feeds are b o i l e d in w a t e r
d u r i n g t h e s t a i n i n g process. F u r t h e r m o r e , t h e
r a r e - e a r t h e l e m e n t s are easily a n a l y z e d b y using
n e u t r o n a c t i v a t i o n analysis. T h e s e m e t h o d s
s h o u l d b e o f help in a n s w e r i n g q u e s t i o n s
p e r t a i n i n g t o rate o f ingesta passage, a n d h o w
such things as level o f i n t a k e , forage t o grain
ratio, associative effects o f feeds, a n d physical
f o r m o f diet i n f l u e n c e ingesta r e t e n t i o n time.
Also, if i n c r e a s e d fluid t u r n o v e r rate in t h e
r u m e n is i n d e e d c o r r e l a t e d w i t h m i c r o b i a l
p r o t e i n p r o d u c t i o n , t h e n t h e s e m e t h o d s will
p r o v i d e t h e m e a n s for isolating t h e animals,
feeds or c o n d i t i o n s w h i c h p r o m o t e g r e a t e r
t u r n o v e r rates.
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