Crude Protein Supplementation To Reduce Lupine Consumption

by Pregnant Cattle in the Scablands of Eastern Washington

USDA-ARS Poisonous Plant Research Laboratory, Logan, Utah, USA

*Corresponding author: James A. Pfister, [email protected]

Abstract

Lupinus leucophyllus (velvet lupine) is prevalent in eastern Washington State, and when
consumed by pregnant cows, it can cause “crooked calf syndrome.” Rangelands in this
region are dominated by poor-quality annual grasses. The objective of this study was to
determine whether feeding supplemental crude protein (CP) would reduce cattle
consumption of velvet lupine during midsummer. Twelve pregnant cows were divided
into 2 treatments: 6 controls received no supplement, and 6 cows received a supplement
of 44% soybean meal at 4 g/kg body weight per day for 19 days in July 2010. Lupine
density was >10 plants/m2. There was no supplement effect on lupine consumption (P =
0.68), nor was there a day x supplement interaction (P = 0.88). Supplemented cattle took
6.9 ± 0.7% of daily bites as lupine compared to 5.0 ± 0.6% of bites for controls. Dry
grass and dry forbs were the major diet components selected by cattle. Cattle ate mostly
dry grass during the first 8 days of the trial, but then their consumption of dry forbs
increased substantially. Cattle switched from eating green forbs to lupine after about 1
week; this change coincided with a decrease in green forb biomass. Five calves from both
treatments were born with mild to moderate birth defects. We conclude that CP
supplementation had no effect on cattle consumption of lupine under these supplement
and abundant-forage conditions. The combined effect of supplementation and drought
(i.e., forage scarcity) on lupine consumption by cattle has not been determined.

Keywords: cattle, lupine, protein supplementation

Introduction

Lupines (Lupinus spp.) are prevalent in the rugged offspring, so-called “crooked calves.” The incidence
Scabland region of eastern Washington State. of crooked calf syndrome on ranches where velvet
Rangelands in this region are dominated by annual lupine is prevalent is estimated to be about 5% per
grasses, including Bromus tectorum (cheatgrass) and year. Some years the incidence is much higher,
Taeniatherum caput-medusae (medusahead), and a typically when lupine populations are very dense
number of weedy forbs, such as Chondrilla juncea from increased winter and spring precipitation
(skeletonweed) and Amsinckia lycopsoides (Panter et al. 2009).
(fiddleneck). Lupines are deep-rooted legumes that Our studies on this type of rangeland indicate
are grazed in mid- to late summer, when annual that grazing cattle prefer graminoids, particularly
grasses are dormant and other forbs have been green grass when available (Lopez-Ortiz et al. 2007,
depleted by grazing. One dominant lupine species is Pfister et al. 2008). However, at times cattle will
Lupinus leucophyllus (velvet lupine), which contains consume relatively large amounts of forbs (>60% of
the teratogenic alkaloid anagyrine. When pregnant the diet) depending on the forage milieu (Lopez-
cows graze velvet lupine between days 40 and 100 Ortiz et al. 2007, Pfister et al. 2008). Some studies
of gestation, they are at risk of having deformed have suggested that cattle may include large
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 IJPPR, vol. 3, Winter 2014

amounts of forbs in their diets because of forbs’ high Committee and were conducted under veterinary
protein content (Pieper and Beck 1980, Holechek supervision. Twelve 2-year-old Hereford x Angus
1984). (479 kg ± 13 kg body weight) cows were time bred
Altering the nutritional status of cattle may so that they were between 40 and 60 days pregnant
influence their forage preference (Heady 1964). at the start of this study. They were verified pregnant
Protein supplementation can influence an animal’s by rectal palpation before the study began and
nutritional status (Kartchner 1980). However, palpated upon return to Logan, UT, shortly after the
protein supplementation has had limited success in end of the study. The 12 cows were divided into 2
influencing botanical diet selection by grazing cattle treatments: 6 controls receiving no supplemental
on grass-forb rangelands (Judkins et al. 1985). In feed and 6 cows receiving a supplement of 44%
contrast, Odadi et al. (2013) reported that protein soybean meal at 4 g/kg body weight per day.
supplementation reduced forb grazing by cattle in Supplemental feeding began 10 days before the
African savanna during the dry season. For grazing trial was started.
ruminants grazing on shrub-dominated rangelands A lupine-infested pasture was subdivided into 6
(e.g., sagebrush), providing supplemental protein smaller pastures such that lupine was abundant in all
and/or energy to detoxify terpenes may greatly pastures, and 2 cows from the same treatment were
enhance shrub use by ruminants (Villalba et al. placed into each of the 3 replicate pastures. Cattle
2006, Dziba et al. 2007). were placed into a corral at night at 1900 h each day,
If protein supplementation could reduce with water and salt provided ad libitum. Supplement
selection of plants such as toxic lupine by grazing was fed to each animal individually in the corral
livestock, then it could be a useful tool to reduce the during the evening. Lupine consumption was
risk of crooked calf syndrome on Scabland determined using bite counts. Each morning at 0530
rangelands. During a typical summer in the h, cattle were released in pairs into their respective
Scablands, all grasses and many forbs become dry pastures to graze, and diets were determined using
and senescent, with resultant decreases in bite counts. Bite counts were taken on individual
concentrations of crude protein, compared to forages animals during active grazing periods; categories for
that remain green later into the summer. Grazing bite counts were lupine (subcategories: whole plant,
cattle typically increase their consumption of lupine leaves, flowers, and pods), dry grass, green grass,
in midsummer (Ralphs et al. 2006). Pregnant heifers dry forbs (other than lupine), and green forbs. Each
require approximately 9% to 10% crude protein (CP) animal was observed for a 5-min period, then the
in their diet to gain 0.75 kg/day (NRC 2000). Our observer moved to the next animal. In this manner,
hypothesis was that cattle would reduce the observer rotated through the entire group of
consumption of still-green lupine in midsummer animals several times each day. Animals were
after the grasses had senesced if the cattle were accustomed to the procedure, and the observer could
supplemented with high-quality protein. Thus, the remain within 1 to 2 m of the animals without
purpose of this trial was to determine whether disturbing their grazing. Typically, about 30 min of
providing a CP supplement to pregnant cattle would daily observation time was recorded for each animal.
alter their selection of velvet lupine during Forage availability and quality were evaluated
midsummer grazing. by weekly clipping of plants to ground level in 10
plots (0.25 m2) within each of the 6 pastures.
Materials and Methods Clipped material was sorted into dry grass, green
grass, lupine, other green forbs, and other dry forbs.
The grazing study was conducted from July 8 to July The clipped material was dried at 40 °C for 48 hours
26, 2010, near Washtucna, WA (46° 48' 12.8" N and weighed to determine available forage
118° 17' 08.5" W; 529 m elevation). Vegetation on (kilograms per hectare). The same plant material was
the study site was dominated by cheatgrass and also retained for nutritional analysis. Lupine density
medusahead; other annual species included was determined at the beginning of the study by
fiddleneck, tumblemustard (Sisymbrium altissimum), placing 150 quadrats, 0.5 m2, along 3 pace transects
tansy mustard (Descurainia pinnata), prickly lettuce that trisected all of the pastures, and counting all
(Lactuca serriola), and black mustard (Brassica lupine plants within each plot.
nigra). Velvet lupine and vetch (Vicia cracca) were Five individual clipped samples of each forage
abundant in all pastures. category for each time point were ground to pass a
All procedures were approved by the Utah State 1-mm screen in a Wiley mill and analyzed for dry
University Institutional Animal Care and Use matter, CP (N x 6.25; LECO FP-528 Nitrogen

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Pfister et al.: Supplementation to reduce lupine consumption

Analyzer, LECO Corp., St. Joseph, MI), neutral standard curve over the range of 25 to 800 µg/mL
detergent fiber (NDF) (ANKOM Fiber Analyzer anagyrine in methanol prepared by serial dilution.
system), and in vitro true digestibility (ANKOM Climatic data before and during the study were
Daisy II system). The NDF procedure was modified taken from a National Climate Data Center weather
by addition of heat-stable amylase (Sigma Chemical, station in Endicott, WA, and a local meteorological
St. Louis, MO). station in Washtucna, WA. Analysis of all data was
The alkaloid composition of lupine (Lee et al. performed using SAS (SAS Institute Inc., Cary,
2007) was determined on 10 randomly selected NC). The bite-count data were checked for normality
whole plants collected weekly. Extraction of L. and transformed using an arcsine square root
leucophyllus was done by weighing 100 mg of transformation typically used for proportions
ground plant material into a 10-mL screw-top test ranging from 0 to 100%. All means shown here are
tube equipped with Teflon-lined caps. Five nontransformed means. The bite-count data were
milliliters of 1N hydrochloric acid (HCl) and 4 mL analyzed using a mixed linear model (Proc Mixed in
chloroform (CHCl3) were added to the test tubes, SAS), with pasture as the experimental unit; animals
which were placed in a mechanical shaker for 15 as a random factor nested within pasture and
min, then centrifuged to separate the aqueous and treatment; treatment; days; and interactions. The
organic phases. The aqueous fraction was transferred probability-of-difference option was used for
to a clean test tube and basified to a pH of between 9 preplanned comparisons.
and 9.5 by drop-wise addition of concentrated
ammonium hydroxide (NH4OH). The basified Results
solution was extracted twice with CHCl3 (4 mL, then
2 mL). The combined CHCl3 fractions were filtered The mean maximum, minimum, and average
through anhydrous sodium sulfate (Na2SO4) and temperatures during the study period were 33, 18,
then dried under a stream of nitrogen (N2) at 50 °C and 25 °C, respectively. There was no precipitation
to yield the crude alkaloid fraction. during the study. Normal January-to-June
The alkaloid fraction extracted from L. precipitation in Washtucna is 14.5 cm (NOAA
leucophyllus was reconstituted in 4 mL of methanol 2002); during 2010, precipitation during this period
containing 1.3 µg/mL caffeine (internal standard). A was 14% above normal.
portion (~1 mL) was transferred to 1.5-mL gas Lupine was abundant and uniformly distributed
chromatograph (GC) autosample vials, and 2 μL across the pastures. Mean (± SE) lupine density for
were injected into an HP 5890 GC (HP 5890, plants of all phenological growth stages and across
Agilent, Palo Alto, CA) equipped with a all 6 small pastures was 10.8 ± 1.0 plants/m2. There
split/splitless injector, FID detector, and a J&W DB- was ample dry grass during the study, but virtually
5 (30 m x 0.33 mm i.d.) capillary column. Injector no green grass in the pastures (table 1). Conversely,
temperature was 250 °C and operated in the splitless there was an abundance of green forbs. Much of the
mode. Split vent flow rate was 60 mL/min and green forb component was V. cracca, which
purged after 1 min. Oven temperature was eventually changed to the dry-forb category, with a
programmed: 100°C for 1 min; 100 to 200°C at few other weedy plants such as rush skeletonweed
50°C/min; and 200 to 320°C at 5°C/min. Caffeine also present.
was used as an internal standard, and plant alkaloid The anagyrine concentration in the lupine
peaks were quantified against a 6-point anagyrine population was near or above 0.2% during the study

Table 1. Forage availability (kg/ha ± SEM) of green and dry grasses and forbs, and lupine (Lupinus
leucophyllus) in eastern Washington during July 2010.
Date Dry Grass Green Grass Dry Forbs Green Forbs Lupine
July 8 701 ± 102 18 ± 9 29 ± 15 524 ± 63 766 ± 126
July 15 497 ± 39 0 85 ± 14 206 ± 37 372 ± 68
July 22 210 ± 33 0 156 ± 27 185 ± 22 281 ± 53

Table 2. Alkaloid concentration (%) of lupine (Lupinus leucophyllus) in eastern Washington

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Figure 1. Nutrient content (% of dry matter) in dry grass, green grass, other forbs, and lupine (Lupinus leucophyllus) in
eastern Washington during July 2010. Crude protein is N x 6.25; NDF = neutral detergent fiber; digestibility = in vitro true
digestibility.

Figure 2. Diet selection (% of bites ± SE) by cattle during July 2010 when receiving either a protein
supplement (soybean meal) or no supplement (controls) while grazing a lupine-infested rangeland in
eastern Washington.

period (table 2). Total alkaloid concentrations slightly above 50%; Digestibility of green forbs and
exceeded 0.85% on the three sampling dates. dry grass was >60% at the start of the study and
Over the study period, the average CP content decreased only slightly during the study period
was 5.3% for dry grass, 6.7% for dry forbs, and (figure 1).
7.7% for green forbs (including vetch). These values Dry grass and dry forbs were the major diet
are in contrast to the average CP content of vetch components during much of the study (figure 2).
alone (8.4%) and lupine (10.5%). Dry grass Cattle ate mostly dry grass during the first 8 days of
contained the highest concentrations of NDF the trial, then dry grass consumption became more
(>70%), whereas lupine and green forbs contained variable (figure 2). There was a day x supplement
less NDF (50–60%). As the lupine matured during interaction (P = 0.04), with control animals eating
the study, digestibility decreased from >60% to more dry grass on days 23 and 24. The consumption

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Pfister et al.: Supplementation to reduce lupine consumption

of forbs such as V. cracca increased substantially in grasses were nearly all senesced when the study
the latter portion of the trial, but there was no began. By the last week of the study, virtually all
treatment effect or treatment x day interaction (P = lupine plants on the study site were at least partially
0.08). There was no treatment (i.e., supplement) dry from the seasonally hot temperatures (>38 °C)
effect on lupine consumption (P = 0.68), nor was and desiccating wind. Under these forage and
there a day x supplement interaction (P = 0.88). climatic conditions, there was no effect of CP
Numerically, supplemented animals ate substantially supplementation on lupine consumption by pregnant
more lupine on several days during the trial (figure cattle.
2), but animal-to-animal variability was high, and no As noted in other studies (Ralphs et al. 2006,
statistical differences were found in lupine Pfister et al. 2008), lupine is not a preferred forage in
consumption between supplemented and control these annual rangelands, but cattle eat lupine after
animals. Overall, supplemented cattle took 6.9 ± the grasses are senesced. Further, in this study, cattle
0.7% of daily bites as lupine compared to 5.0 ± 0.6% increased lupine consumption and greatly
of bites for control cattle. Interestingly, cattle began diminished consumption of green forbs about
to switch from green forbs to lupine in their diets midway through the study period. This dietary
after about 1 week on the study pastures, and this switch coincided with increased consumption of dry
switch coincided with a decrease in available green forbs, particularly vetch, the majority of which
forb biomass about midway through the trial. senesced during this period on the study site. Lopez-
One pregnant cow resorbed her fetus during the Ortiz et al. (2007) and Ralphs et al. (2011) also
grazing study. Five other calves were born with mild found that cattle selected lupine only after
to moderate arthrogryposis, scoliosis, and/or availability of annual grasses and other forbs
rotational defects (i.e., impaired leg movement) in declined. Our observations indicate that cattle
the front legs (table 3). No cleft palates were noted. typically began to graze lupine plants near the top,
There was no apparent relationship between eating floral parts, pods, leaves, and stem, then often
supplementation and the occurrence of birth defects. regrazed the same plants several times, eventually
eating the plants down to short, bare stalks.
Table 3. Deformities in calves born to pregnant cows
One can speculate that the cattle may have
which grazed on lupine-infested pasture from July 8 to responded differently to the supplement if lower
26, 2010, within the gestational day 40 to 70 window precipitation had resulted in reduced biomass for
for birth defects. + signifies slight defects, and ++ other forbs, particularly the relatively palatable
indicates moderate defects. S= Supplement; vetch. The abundant vetch was consumed when
C=Controls. No cleft palates were noted. green, and cattle continued to consume the vetch
Deformity after it senesced. However, our previous experience
Cow Treat- Arthro- Scoliosis Leg in this region suggests that lupine populations are
# ment gryposis movement
62 S + +
also depressed during drought (Pfister et al. 2008).
59 S + Lupine density at this site was >10 plants/m2, which
61 S + contrasts greatly with lupine density measured at
36 C ++ another nearby site during a moderate drought (1-2
57 C ++ +
plants/m2; Pfister et al. 2008). The effectiveness of
any supplementation regime to reduce lupine
consumption by cattle during drought in the
Discussion
Scablands region is not known. However, recurring
drought conditions in the Scablands region since
There was ample plant biomass for cattle diet
1997 have reduced lupine populations and
selection throughout the entire study period; near the
diminished the incidence of crooked calf syndrome
end of the study, there were declining but still-
(Panter et al. 2009). Gay et al. (2007) also noted an
sufficient amounts of green and dry forbs, dry grass,
association between lupine density in pastures and
and lupine across the pastures. It was apparent that
the reported incidence of “crooked calves.”
the vegetation, including lupine and other forbs,
It is interesting to note that some “weedy” forbs
responded favorably in terms of biomass to the
such as rush skeletonweed are relatively palatable
wetter-than-normal winter and spring conditions.
for cattle on these degraded annual rangelands; the
This site was selected specifically because of the
stemmy skeletonweed, for example, is listed as a
abundant lupine population during some years.
noxious weed in eastern Washington and elsewhere.
Because of the intentional timing of the study, the
However, at times cattle consumed relatively large
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 IJPPR, vol. 3, Winter 2014

quantities of this weed. On these annual rangelands, anagyrine dose and duration necessary to cause
cattle also consumed large, but declining, quantities defects.
of cheatgrass and also some of the very coarse We conclude that supplemental protein had no
medusahead. We did not distinguish between effect on lupine consumption by pregnant cattle.
consumption of the various annual grasses in our This study was conducted under conditions of
bite counts, but in the absence of other, more abundant forage, including lupine and other forbs,
palatable green grasses, cattle readily accepted the and these conditions undoubtedly had a major
dry annual grasses, primarily cheatgrass, as forage. impact on the findings. With normal precipitation
The CP content of lupine and vetch was patterns in conjunction with overgrazing, it is
numerically higher than other available forages such possible that the depletion of forbs by grazing may
as dry grass or dry forbs. However, it is important to not go hand in hand with lupine depletion, and under
note that these CP concentrations are taken from this scenario the likelihood of a positive effect from
clipped plant material, and numerous studies have protein supplementation may increase. It is unknown
shown that cattle can select a diet substantially whether cattle grazing on lupine-infested rangelands
higher in nutritional composition than is shown in during drought would alter their lupine consumption
available plant material by clipping (Weir and Torell if given a CP supplement, because lupine
1959, Galt et al. 1969, Coleman and Barth 1973). populations also decline during drought along with
That was undoubtedly true for cattle in this study as other forage classes.
well. Thus, we speculate that even unsupplemented
cattle in this study were able to meet their minimal Acknowledgments
nutritional needs for CP (about 9% CP in diet)
without grazing toxic lupine on this annual grassland We thank Kermit Price for assistance with all
site during midsummer.Soybean meal is a rich aspects of the study from laboratory to the field,
source of protein, with both rumen degradable Chuck Hailes for anagyrine analysis, and Rex Probst
protein (RDP, 65% of CP) and rumen undegradable for animal care and transportation during the study.
protein (RUP, 35% of CP). Many forage grasses
contain mostly RDP (Buckner et al. 2013), at levels References
similar to those in soybean meal. There is no
information available on the relative RDP and RUP Buckner CD, Klopfenstein TJ, Rolfe KM, et al. 2013.
concentrations of Scabland forbs or lupine. It is Ruminally undegradable protein content and digestibility
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0.20% found in this study are either similar or Animal Science 36:754-761.
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this area for whole plant material (Ralphs et al. 2006, Supplemental energy and protein increase use of
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mild to moderate, suggesting that the lupine central Washington state. In KE Panter, TL Wierenga, JA
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