Effect of an injectable trace mineral at the initiation of a 14 day

CIDR protocol on heifer performance and reproduction1

R. S. Stokes, A. R. Ralph, A. J. Mickna, W. P. Chapple, A. R. Schroeder, F. A. Ireland, and D. W. Shike2
Department of Animal Sciences, University of Illinois, Urbana 61801

ABSTRACT: Three experiments were conducted at AI and overall pregnancy rates were also similar (P ≥
separate locations to determine the effects of a trace 0.74) regardless of treatment. During Exp. 2, BCS and
mineral injection (TMI), Multimin 90, on heifer perfor- BW did not differ (P ≥ 0.44) across treatments. There

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mance and reproduction. In Exp. 1, [spring-born, Angus, was a tendency (P = 0.07) for TMI heifers to have an
n = 93, body weight (BW) = 428 ± 45.2 kg], Exp. 2 increased AI pregnancy rate (62 vs. 45%) compared
(spring-born, Angus × Simmental, n = 120, BW = 426 ± with control heifers despite no difference (P = 0.51) in
54.0 kg), and Exp. 3 (fall-born, commercial Angus, n = overall pregnancy rate. In Exp. 3, BW was not differ-
199, BW = 345 ± 39.7 kg) heifers were stratified by BW ent (P ≥ 0.39) across all time points. Also, BCS did not
within experiment and assigned to 1 of 2 treatments: a differ (P ≥ 0.45) at initiation, AI, or final pregnancy con-
control, saline injection, or TMI at a dose of 1 mL/68 kg formation. Interestingly, there was a tendency (P = 0.10)
BW. Free choice mineral, containing Cu, Mn, Se, and Zn for TMI heifers to have an increased BCS at the time of
formulated to meet or exceed NRC recommendations, breeding compared with control heifers. However, there
was supplemented to heifers. Injections were given 33 were no differences (P ≥ 0.50) in AI and overall preg-
d prior to breeding at the initiation of a 14-d controlled nancy rates. In 1 of 3 experiments, an injectable trace
internal drug release (CIDR)-prostaglandin protocol. mineral administered 33 d prior to the breeding sea-
There was no difference (P ≥ 0.37) in BW during Exp. son in conjunction with a 14-d CIDR protocol, tended
1. Additionally, there was no difference (P ≥ 0.52) in to increased AI conception rates of heifers even when
body condition score (BCS) at initiation or at artificial adequate trace mineral supplement was provided. The
insemination (AI) and final pregnancy confirmation in variable response observed across experiments may be
Exp. 1; however, a greater (P = 0.03) BCS was noted caused by differences in breed, calving season, mineral
for control heifers at breeding. Pregnancy rates to timed sources, and management strategies.
Key words: artificial insemination, beef heifer, injectable trace mineral, pregnancy rate, reproduction

© 2017 American Society of Animal Science. This is an open access article distributed under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Transl. Anim. Sci. 2017.1:458–466
doi:10.2527/tas2017.0050

INTRODUCTION

Trace minerals such as copper, manganese, sele-

1The authors would like to thank the Illinois Beef Association nium, and zinc play critical roles in biochemical pro-
for partial funding for this project and Multimin USA for donating cesses and are key components of a ruminant animal’s
the Multimin 90 and partial funding for this project. The authors health and productivity (Suttle, 2010). Grazing cattle
would also like to thank the staff at the University of Illinois Beef primarily receive trace minerals through forages; how-
Cattle and Sheep Field Laboratory, Urbana, IL, Orr Agricultural ever, these sources often do not meet cattle require-
Research and Demonstration Center in Baylis, IL, and Dixon ments due to variation in soil composition (Smart et
Springs Agricultural Center, Simpson, IL for care of the experi- al., 1981). In these instances, producers commonly
mental animals and aiding in collection of data. supplement through free-choice mineral, salt blocks
2Corresponding author: [email protected]
fortified with trace minerals, or protein/energy supple-
Received August 1, 2017.
ments fortified with trace minerals (Arthington et al.,
Accepted September 13, 2017.
458
 Injectable trace mineral heifer reproduction 459

2014). Bioavailability of trace mineral sources can vary an injection with sterilized saline (CON1) or an injection
due to interactions with other minerals and feed com- of Multimin 90 (MM1) administered subcutaneously at
ponents within the gastrointestinal tract (Spears, 2003). a dose of 1 mL/68 kg BW. The Multimin 90 contained
An injectable trace mineral provides the opportunity to 60 mg/mL of zinc as zinc oxide, 10 mg/mL of manga-
supplement trace minerals that completely bypass the nese as manganese carbonate, 5 mg/mL of selenium as
gastrointestinal tract and thus avoid the complex ruminal sodium selenite, and 15 mg/mL of copper as copper car-
interactions. Also, a multi-element injectable trace min- bonate. Prior to the start of the trial, heifers were weaned
eral allows for targeted delivery of a specific amount to and developed on a diet consisting of roughage, corn co-
individual animals (Arthington et al., 2014) and elimi- products, and supplement. Heifers were then adapted to
nates the variability associated with voluntary intake of a total mixed ration (TMR, Table 1) that included both
free choice mineral (Arthington and Swenson, 2004). inorganic and organic trace mineral, and remained on
Pogge et al. (2012) recently demonstrated trace the same diet through the initiation of the trial. In year 1
mineral injections are an effective way to increase the heifer initial BW was collected on d 0 (April 2015) and

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trace mineral status of calves, particularly Cu and Se. heifers were enrolled in a 14-d CIDR (Pfizer Animal
Increased mineral status may be of particular impor- Health, New York, NY) insert – PG and timed AI proto-
tance when biological needs are increased, such as col (Mallory et al., 2012). On d 19 heifers were transport-
breeding. In some research, an injectable trace mineral ed to pasture where they grazed 70% endophyte-infected
has improved reproductive performance. Sales et al. fescue (Festuca arundinacea), and 30% red (Trifolium
(2011) reported injectable trace mineral supplementa- pretense) and white clover (Trifolium repens) pastures
tion increased conception rates and chance of embryo (60.91% NDF, 32.28% ADF, and 13.04% CP). While on
survival after timed embryo transfer. Additionally, pasture, heifers had access to free choice mineral [19.7%
Kirchhoff (2015) reported an injectable trace mineral
increased heifer artificial insemination (AI) concep-
tion rate. Multimin 90 is labeled for administration 30 Table 1. Ingredient composition of heifer diets (% DM
d prior to breeding, which coincides with the initia- basis) for Exp. 1
tion of a 14-d controlled internal drug release (CIDR) Inclusion, % DM
prostaglandin (PG) protocol. From a practical man- Item Year 1 Year 2
agement standpoint, this allows for the administration Ingredient, %
of injectable trace mineral without any additional han- Corn silage 72 60
dling. Therefore, the objective of these experiments MDGS1 13 15
was to assess the effects of an injectable trace mineral Treated corn stalks2 10 –
Grass hay – 20
administered at the initiation of a 14-d CIDR protocol
Supplement3,4 5 5
on heifer performance and reproduction.
Analyzed nutrient content
CP, % 10.1 9.9
MATERIALS AND METHODS NDF, % 51.1 50.7
ADF, % 28.9 27.7
All experimental procedures were approved by the Crude fat, % 2.9 3.5
Institutional Animal Care and Use Committee of the S, % 0.18 0.20
University of Illinois (IACUC #16046) and followed Cu, mg/kg 4.8 13.6
the guidelines recommended in the Guide for the Mn, mg/kg 29.3 29.1
Care and Use of Agricultural Animal in Agricultural Zn, mg/kg 21.9 36.6
Research and Teaching (FASS, 2010). 1Experiment 1 determined the effects of a TMI on spring born Angus ×
Simmental heifer performance and reproduction.
2Modified distillers grains with solubles.
Experiment 1 3Corn stalks were treated with Silage SAVOR Plus (Kemin Industries,
Inc., Des Moines, IA) at 0.5 kg Mg–1 applied at bagging.
Animals and Experimental Design. Spring-born, 4Supplement contained 87.7% ground corn, 8.9% limestone, 1.8% trace
Angus heifers [Year 1: initial body weight (BW) = 430 mineral salt [8.5% Ca as calcium carbonate, 5% Mg as magnesium oxide and
± 489.9 kg, n = 40; Year 2: initial BW = 425 ± 40.5 kg, n magnesium sulfate, 7.6% K as potassium chloride, 6.7% Cl as potassium chlo-
= 53] housed at the University of Illinois Beef Cattle and ride, 10% S as S8, prilled, 0.5% Cu as copper sulfate and Availa-4 (Zinpro
Performance Minerals; Zinpro Corp, Eden Prairie, MN), 2% Fe as iron sulfate,
Sheep Field Laboratory in Urbana, IL were utilized to as- 3% Mn as manganese sulfate and Availa-4, 3% Zn as zinc sulfate and Availa-4,
sess the effects of an injectable trace mineral (Multimin 278 mg/kg Co as Availa-4, 250 mg/kg I as calcium iodate, 150 mg/kg Se as
90, Multimin USA, Fort Collins, CO) on heifer reproduc- sodium selenite, 2,205 KIU/kg VitA as retinyl acetate, 662.5 KIU/kg VitD as
cholecalciferol, 22,047.5 IU/kg VitE as DL-α-tocopheryl acetate, and less than
tion and performance over a 2-yr period. Heifers were 1% crude protein, fat, crude fiber, salt], 0.1% Rumensin 90 (198 g monensin/
stratified by BW and assigned to 1 of 2 treatments: either kg, Rumensin 90; Elanco Animal Health, Greenfield, IN), and 1.5% fat.
Translate basic science to industry innovation
460 Stokes et al.

limestone, 19.7% trace mineral salt (8.5% Ca as calcium 1988; The Ohio State University, Service Testing and
carbonate, 5% Mg as magnesium oxide and magnesium Research Lab, Wooster, OH).
sulfate, 7.6% K as potassium chloride, 6.7% Cl as potas- Statistical Analysis. Body weight and BCS were
sium chloride, 10% S as S8, prilled, 0.5% Cu as cop- analyzed using the MIXED procedure of SAS (SAS
per sulfate and Availa-4 (Zinpro Performance Minerals; Inst. Inc., Cary, NC). Artificial insemination rates
Zinpro Corp, Eden Prairie, MN), 2% Fe as iron sulfate, and overall pregnancy rates were analyzed using the
3% Mn as manganese sulfate and Availa-4, 3% Zn as GLIMMIX procedure of SAS. Final pregnancy confir-
zinc sulfate and Availa-4, 278 mg/kg Co as Availa-4, 250 mation data included bull in the model; however, this
mg/kg I as calcium iodate, 150 mg/kg Se as sodium sel- was not significant (P = 0.57), so bull was removed
enite, 2,205 KIU/kg VitA as retinyl acetate, 662.5 KIU/ from the model. Heifer was considered the experi-
kg VitD as cholecalciferol, 22,047.5 IU/kg VitE as DL- mental unit for all measures. The model included the
α-tocopheryl acetate, and less than 1% crude protein, fat, fixed effect of treatment and the random effect of year.
crude fiber, salt), 14.2% salt, 40.6% monocalcium phos- Significance was declared at P ≤ 0.05, and tendencies

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phate, 4.2% dried molasses, 1.4% zinc sulfate, and 0.01% were declared from 0.05 < P ≤ 0.10. Means reported in
iodine]. Heifers were weighed, body condition scored tables are least squares means ± SEM.
(BCS), and AI on d 33. Ten d following AI, heifers were
exposed to 1 bull that had previously passed a breeding Experiment 2
soundness exam, for a 64 d breeding season. Artificial
insemination conception rates, BCS, and BW were col- Animals and Experimental Design. To determine
lected on d 70 and final BCS, weights, and overall preg- the effects of an injectable trace mineral on heifer re-
nancy confirmation were collected on d 130. A trained production and performance over a 2-yr period, spring-
technician determined artificial insemination conception born, Angus × Simmental heifers (Year 1: initial BW
and overall pregnancy rates via ultrasonography (Aloka = 411 ± 48.1 kg, n = 65; Year 2: initial BW = 441 ±
500, Hitachi Aloka Medical America, Inc., Wallingford, 59.8 kg, n = 55) were utilized. Heifers were either 0.5
CT; 7.5 MHz general purpose transducer array). Angus and 0.5 Simmental or 0.625 Angus and 0.375
In year 2, heifers were managed similarly with Simmental. Cattle were stratified by BW into 2 groups
the following exceptions. At the initiation of the trial and randomly assigned either a sterilized saline injec-
heifer BCS was measured. Ten d following AI heifers tion (CON2) or a Multimin 90 injection (MM2) at a
were divided into 2 groups with equal treatment rep- rate of 1 mL/68 kg BW. In yr 1, prior to the start of the
resentation in each group and exposed to 1 bull/group trial, heifers were housed at the University of Illinois
that had previously passed a breeding soundness exam. Beef Cattle and Sheep field Laboratory in Urbana, IL
On d 60 (17 d after heifers had been exposed to bulls) and fed a diet consisting of alfalfa haylage, corn si-
1 bull had to be removed from the study, so heifers lage, corn co-products, and supplement. Heifers were
were again co-mingled and placed with 1 bull that had then adapted to a TMR (Table 1) and remained on this
passed a breeding soundness exam. Final BCS, BW, diet until d –13, which included both an organic and
and pregnancy confirmation were collected on d 145. inorganic trace mineral supplement. On d –13 heif-
Sample Collection and Analytical Procedures. ers were transported to the Orr Agricultural Research
For nutrient composition analysis, individual feed and Demonstration Center in Baylis, IL. Heifers were
ingredients were collected monthly and composited housed in soil surface pens and offered a TMR and an
and dried at 55°C for a minimum of 3 d and ground inorganic trace mineral supplement (Table 2). Intake
through a 1-mm screen using a Wiley mill (Arthur, H. averaged 7.1 kg of DM/d per heifer across treatments.
Thomas, Philadelphia, PA). Forage samples were col- At the initiation of the study (April, 2015), individual
lected from pastures on a monthly basis and compos- heifer BW were measured, treatments applied, and
ited and dried similarly. Ground feed samples were heifers were enrolled in a 14-d CIDR-PG and timed
analyzed for CP (Leco TruMac, LECO Corporation, AI protocol. On d 33 heifers were weighed, BCS,
St. Joseph, MI), NDF and ADF using an Ankom and timed AI. Immediately following timed AI heif-
200 Fiber Analyzer (Ankom Technology, Macedon, ers were transported to pasture for the duration of the
NY), and crude fat using an Ankom XT10 fat extrac- breeding season. Heifers grazed pastures with an aver-
tor (Ankom Technology). Ground forage samples age coverage area of 70% endophyte-infected fescue
were also analyzed for NDF, ADF, and CP. Forage (Festuca arundinacea) and 30% red clover (Trifolium
and feed samples were also sent to a commercial lab pretense) and white clover (Trifolium repens, 9.05%
where they were subjected to nitric acid digestion NDF, 32.98% ADF, and 11.96% CP) for the remain-
and inductively coupled plasma spectroscopy analy- der of the study and were given access to free choice
sis for complete minerals (method 975.03: AOAC, mineral [12% Ca as calcium carbonate, 8% P, 18%
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 Injectable trace mineral heifer reproduction 461

Table 2. Ingredient composition of heifer diets (% DM were housed in soil surface pens and received a similar
basis) for Exp. 21 diet and trace mineral supplement (Table 2). Average
Inclusion, % DM heifer DMI was 8.5 kg/d per heifer across treatments.
Item Year 1 Year 2 At the time of CIDR removal, all heifers’ tail heads
Ingredient, % were painted and paint scores (1 = completely gone, 2
Corn silage 24.7 59.0 = partially gone, and 3 = untouched) were collected at
MDGS2 14.14 20.0 the time of breeding. Heifers were managed in 2 groups
Grass hay 61.16 16.0
and 10 d following AI were exposed for a 46 d breeding
Supplement3,4 – 5.0
season to 1 bull per group that had previously passed a
Analyzed nutrient content
breeding soundness exam. Overall pregnancy rate, BW,
CP, % 12.1 10.5
NDF, % 53.9 41.1
and BCS were collected on d 145.
ADF, % 33.2 21.0 Sample Collection and Analytical Procedures.
Crude fat, % 2.3 4.2 Feed ingredients and forage samples were collected

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S, % 0.27 0.22 and analyzed as described in Exp. 1.
Cu, mg/kg 6.4 7.0 Statistical Analysis. Data were analyzed as de-
Mn, mg/kg 40.2 67.2 scribed in Exp. 1 with the following exception that
Zn, mg/kg 30.0 37.8 group was included as a fixed effect. Treatment dis-
1Experiment 2 determined the effects of a TMI on spring born Angus × tributions of tail paint scores were determined using
Simmental heifer performance and reproduction. PROC GLIMMIX of SAS.
2Modified distillers grains with solubles.
3In year 1 supplement was top dressed at a rate of 0.11 kg/heifer per Experiment 3
d (23.4% Ca as calcium carbonate, 15.7% salt, 1.0% Mg as magnesium
oxide, 3,500 mg/kg of Zn as zinc sulfate, 3,350 mg/kg of Cu as copper
sulfate, 26.4 mg/kg of Se as sodium selenite, 181,437 IU/kg of vitamin A, Animals and Experimental Design. Fall-born,
and 181 IU/kg of vitamin E). commercial Angus (n = 199, initial BW = 345 ± 39.7
4Supplement contained 87.7% ground corn, 8.9% limestone, 1.8% trace
kg) were utilized to determine the effects of an inject-
mineral salt [8.5% Ca as calcium carbonate, 5% Mg as magnesium oxide and
magnesium sulfate, 7.6% K as potassium chloride, 6.7% Cl as potassium chlo- able trace mineral on heifer BW, BCS, and repro-
ride, 10% S as S8, prilled, 0.5% Cu as copper sulfate and Availa-4 (Zinpro ductive performance. Heifers were fed and managed
Performance Minerals; Zinpro Corp, Eden Prairie, MN), 2% Fe as iron sulfate, at the Dixon Springs Agricultural Research Center,
3% Mn as manganese sulfate and Availa-4, 3% Zn as zinc sulfate and Availa-4,
278 mg/kg Co as Availa-4, 250 mg/kg I as calcium iodate, 150 mg/kg Se as
Simpson, IL. Body weight and sire were used to strat-
sodium selenite, 2,205 KIU/kg VitA as retinyl acetate, 662.5 KIU/kg VitD as ify cattle. Heifers were utilized previously in another
cholecalciferol, 22,047.5 IU/kg VitE as DL-α-tocopheryl acetate, and less than study and thus heifers were also stratified by previ-
1% crude protein, fat, crude fiber, salt], 0.1% Rumensin 90 (198 g monensin/
ous treatment (Kordas et al., 2017). On d 0 (October,
kg, Rumensin 90; Elanco Animal Health, Greenfield, IN), and 1.5% fat.
2015), initial BW and BCS were collected, heifers
were synchronized with a 14-d CIDR-PG timed AI
salt, 11% Mg as magnesium sulfate, 90 mg/kg I as protocol as previously described by Mallory et al.
calcium iodate, 108,862 IU/kg vitamin A, 18,144 IU/ (2012), and administered 1 of 2 treatments: a control
kg vitamin D3, 454 IU/kg vitamin E, and 5,600 mg/ sterile saline injection (CON3) or a Multimin 90 in-
kg of chlortetracycline (Aureomycin; Alpharma Inc. jection (MM3) both administered at a rate of 1 mL/68
Animal Health, Bridgewater, NJ)]. Ten days follow- kg of BW. Heifers were placed on pastures following
ing AI, a bull was placed with each pen of heifers for AI, managed as a single group, and grazed 70% en-
a 45 d breeding season. Artificial insemination con- dophyte infected fescue (Festuca arundinacea) and
ception rates, BW, and BCS were determined on d 70. 30% red clover (Tri-folium pretense) pastures (60.53%
Overall pregnancy rate, BW, and BCS were collected NDF, 34.41% ADF, and 9.81% CP). Heifers were of-
on d 144. A trained technician determined artificial fered a supplement consisting of 50% soybean hulls
insemination conception and overall pregnancy rates (62.53% NDF, 45.74% ADF, and 11.26% CP) and
via ultrasonography (Aloka 500 instrument, Hitachi 50% corn gluten feed pellets (33.65% NDF, 9.19%
Aloka Medical America, Inc., Wallingford, CT; 7.5 ADF, and 23.77% CP) at a rate of 2.7 kg/heifer per
MHz general purpose transducer array). d. Free choice loose mineral [Renaissance Nutrition,
In year 2, heifers were managed similarly with the Roaring Springs, PA; 0.16% S, 17.88% Ca as calci-
following exceptions. The University of Illinois Beef um carbonate, 2.99% P as monocalcium phosphate,
Cattle and Sheep field Laboratory in Urbana, IL was 24.5% salt, 9.35% Na, 5.74% Mg as magnesium ox-
used to house heifers from weaning until d 10 of the trial. ide, 0.06% K, 2,214 mg/kg Fe as iron oxide, 2,013 mg/
On d 10, heifers were transported to the Orr Agricultural kg Mn as manganese hydroxychloride (Intellibond M,
Research and Demonstration Center in Baylis, IL and Micronutrients Inc., Indianapolis, IN), 2511 mg/kg Zn
Translate basic science to industry innovation
462 Stokes et al.

as hydroxyl zinc (Intellibond Z, Micronutrients Inc.), Table 3. Influence of an injectable trace mineral on
1,001 mg/kg Cu as tribasic copper chloride (Intellibond heifer BW and BCS over 2 consecutive years in Exp. 1
C, Micronutrients Inc.), 27 mg/kg Co as cobalt car- Treatment1
bonate, 36 mg/kg I as calcium iodate, 26 mg/kg Se as Item CON1 MM1 SEM P-value
sodium selenite, 110,178 IU/kg vitamin A, 3,084 IU/ n, heifer 47 46
kg vitamin D, 545 IU/kg vitamin E, and 1,179 mg/kg BW, kg
of chlortetracycline] was offered ad libitum to heifers. Initial 429 426 4.6 0.72
Breeding 425 424 6.4 0.92
On d 30, heat detection patches (Estrotect Heat
AI pregnancy confirmation 433 418 6.5 0.37
Detectors, Rockway Inc., Spring Valley, WI) were
Final pregnancy confirmation 437 436 16.2 0.78
placed on all heifers. On d 33 heifers were weighed,
BCS
BCS, AI and heat patches were visually scored from Initial 5.4 5.4 0.07 0.52
0 to 3 (0 = missing, 1 = fully activated, 2 = partially Breeding 5.7 5.5 0.17 0.03
activated, and 3 = not activated). Nine days follow- AI pregnancy confirmation 5.5 5.5 0.22 0.86

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ing AI, heifers were exposed to 5 yearling bulls that Final pregnancy confirmation 5.5 5.6 0.22 0.64
had all passed breeding soundness exams, for a 71 d 1Control(CON1) cattle received a sterilized saline solution at 1 mL/68
breeding season. Due to limited pasture forage avail- kg BW, and Multimin 90 (MM1) cattle received injectable trace mineral
ability, on d 71 heifers were offered free choice grass at 1 mL/68 kg BW.
hay (68.7% NDF, 39.9% ADF, and 6.01% CP) for the
remainder of the trial. On d 76 individual BW, BCS,
and AI pregnancy conception rates were collected.
Overall conception rates and final BW and BCS were
collected on d 153. A trained technician determined
AI conception and overall pregnancy rates via ultra-
sonography (Aloka 500 instrument, Hitachi Aloka
Medical America, Inc., Wallingford, CT; 7.5 MHz
general purpose transducer array).
Sample Collection and Analytical Procedures.
Feed ingredients and forage samples were collected
and analyzed as described in Exp. 1.
Statistical Analysis. Heifer BW and BCS were
analyzed using the MIXED procedure of SAS (SAS
Inst. Inc., Cary, NC). Heifer AI conception and overall Figure 1. The effect of an injectable trace mineral (Multimin 90,
pregnancy rates were analyzed using the GLIMMIX MM1) or sterilized saline solution (CON1) at the initiation of a synchroni-
procedure of SAS. Treatment distributions of heat zation protocol on heifer AI and final pregnancy rates over 2 consecutive
years in Exp. 1. Pregnancy rates to timed AI and overall pregnancy rates
patch scores were determined using PROC GLIMMIX were similar for CON1 and MM1 treated heifers (P ≥ 0.74).
of SAS. The model included the fixed effects of treat-
ment. Treatment effects were considered significant at
Experiment 2
P ≤ 0.05 and tendencies were noted at 0.05 < P ≤ 0.10.
All heifers received the same diet (Table 2) or
RESULTS pasture regardless of treatment. Heifer BW and BCS
did not differ across all time points (P ≥ 0.44, Table
4). Additionally, the distribution of tail paint scores
Experiment 1 was similar (P ≥ 0.90) across treatments. There was a
All heifers received the same diet (Table 1) or pas- tendency (P = 0.07, Fig. 2) for MM2 heifers to have
ture regardless of treatment. There was no difference greater AI pregnancy rates (62.1%) compared with
(P ≥ 0.37, Table 3) for heifer BW at any time point. their CON2 (45.2%) counterparts. However, by the
Also, there was no difference (P ≥ 0.52) for heifer time of overall pregnancy confirmation there was no
BCS at the initiation of the trial, AI pregnancy con- difference (P = 0.51) for pregnancy rates regardless of
firmation, or final pregnancy confirmation regardless treatment and overall pregnancy rate averaged 82.7%.
of treatment. Interestingly, CON1 heifers had greater
(P = 0.03) BCS at breeding than their MM1 treated Experiment 3
counterparts. Pregnancy rates to timed AI and over-
all pregnancy rates were similar for CON1 and MM1 Heifer BW did not differ across all time points (P
treated heifers (P ≥ 0.74, Fig. 1). ≥ 0.39, Table 5). Additionally, heifer BCS did not dif-
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 Injectable trace mineral heifer reproduction 463

Table 4. Influence of an injectable trace mineral on Table 5. Influence of an injectable trace mineral on
heifer BW, BCS, and tail paint score over 2 consecutive heifer BW, BCS, and heat patch scores in Exp. 3
years in Exp. 2 Treatment1
Treatment1 Item CON3 MM3 SEM P-value
Item CON2 MM2 SEM P-value n, heifer 99 100
n, heifer 60 60 BW, kg
BW, kg Initial 344 344 2.8 0.93
Initial 427 426 15.1 0.97 Breeding 347 348 2.9 0.79
Breeding 413 413 10.3 0.96 AI pregnancy confirmation 346 348 3.1 0.76
AI pregnancy confirmation 432 427 9.3 0.44 Final pregnancy confirmation 374 370 3.3 0.39
Final pregnancy confirmation 434 435 27.5 0.81 BCS
BCS Initial 5.4 5.4 0.05 0.95
Initial 5.5 5.5 0.09 0.58 Breeding 5.6 5.7 0.05 0.10
Breeding 5.8 5.8 0.09 0.78 AI pregnancy confirmation 5.0 5.1 0.05 0.45

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AI pregnancy confirmation 5.6 5.6 0.14 0.80 Final pregnancy confirmation 4.9 4.9 0.05 1.00
Final pregnancy confirmation 5.5 5.5 0.17 0.58 Heat Patch Score2, %
Tail paint score2,3, % 1 58 59 – 0.89
1 25 23 – 0.90 2 17 20 – 0.59
2 75 74 – 0.95 3 25 21 – 0.50
3 – – – – 1Control (CON3) cattle received a sterilized saline solution at 1 mL/68
1Control (CON2) cattle received a sterilized saline solution at 1 mL/68 kg BW, and Multimin 90 (MM3) cattle received injectable trace mineral
kg BW, and Multimin 90 (MM2) cattle received injectable trace mineral at 1 mL/68 kg BW.
2 Heat patches were visually scored at time of breeding from 0– 3 (0 =
at 1 mL/68 kg BW.
2Tail paint scores were visually assessed at the time of breeding (1 = missing, 1 = fully activated, 2 = partially activated, 3 = not activated). No
completely gone, 2 = partially gone, 3 = untouched). heifers were missing heat patches at the time of breeding.
3Tail paint scores were not collected in year 1.

Figure 3. The effect of an injectable trace mineral (Multimin 90,

Figure 2. The effect of an injectable trace mineral (Multimin 90, MM3) or sterilized saline solution (CON3) at the initiation of a synchroni-
MM2) or sterilized saline solution (CON2) at the initiation of a synchroni- zation protocol on heifer AI and final pregnancy rates in Exp. 3. Pregnancy
zation protocol on heifer AI and final pregnancy rates over 2 consecutive rates to timed AI and overall pregnancy rates were also similar for both
years in Exp. 2. * indicates a tendency (P = 0.07) for MM2 AI conception CON3 and MM3 treated heifers (P ≥ 0.50).
rates to be greater than CON2 heifers. Overall pregnancy rates were simi-
lar for CON2 and MM2 treated heifers (P = 0.51).

DISCUSSION
fer (P ≥ 0.45) at the initiation of the trail or at AI and
final pregnancy confirmation. However, there was a For all experiments, trace mineral injection did not
tendency (P = 0.10) for MM3 heifers to have a greater affect heifer BW at any time point. Also, heifer BCS
BCS at the time of breeding than their CON3 counter- was similar at initiation, AI pregnancy confirmation, and
parts. There were also no differences (P ≥ 0.50) across final pregnancy confirmation regardless of treatment
treatments, in the distribution of heat patch scores across these experiments. Since diet and pasture avail-
for Exp. 3. Pregnancy rates to timed AI and overall ability were the same for both treatments, regardless of
pregnancy rates were also similar for both CON3 and experiment, it is likely this did not impact the results.
MM3 treated heifers (P ≥ 0.50, Fig. 3). These results are consistent with Gadberry and Baldridge
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464 Stokes et al.

(2013), where BW and BCS did not differ for Angus kg average mature BW. The NRC (2016) suggests Bos
cows receiving either an injectable trace mineral or no Taurus heifers should reach puberty at approximately
injectable trace mineral prior to breeding. However, the 60% of mature weight. Additionally, Vera et al. (1993)
effects of injectable trace mineral supplementation on noted heifers in a nutritionally restrictive environment
cow BW and BCS has been inconsistent across literature. will have an even greater percentage of mature BW at
Arthington et al. (2014) reported in beef heifers, those puberty than noted above. Mature cows also in a restric-
administered an injectable trace mineral tended to have tive environment often weigh less than cows of a simi-
greater ADG compared with heifers given saline. Still, it lar genotype not maintained in a restrictive environment
is important to note the heifers utilized by Arthington et (Pahnish et al., 1983). These heifers were more nutri-
al. (2014) were administered treatments at weaning and tionally challenged and thus below their expected per-
heifers remained on study through breeding, for a total cent of mature BW at breeding. The authors speculated
of 177 d. In the current experiments, treatments were ad- the greatest response in trace mineral supplementation
ministered 33 d prior to breeding, at estrus synchroniza- would be noted in Exp. 3. However, pregnancy rates to

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tion, and heifers remained on study until the time of final timed AI and overall pregnancy rates were similar across
pregnancy confirmation, making it challenging to draw treatments, and there were no differences in the distribu-
comparisons across these experiments. tion of heat patch scores for Exp. 3. This data suggests
Interestingly, in Exp. 1, CON1 heifers tended to all heifers responded similarly to estrus synchronization
have a greater BCS at the time of breeding than their and there was no difference in the number of heifers ex-
MM1 counterparts. Contrastingly in Exp. 3, CON3 hibiting standing estrus at the time of AI. This is com-
heifers tended to have a lesser BCS than MM3 heifers parable to work by Brasche et al. (2015) who reported
at the time of breeding. It is important to note these dif- no effect of trace mineral injection on overall pregnancy
ferences in BCS were significant due to a small standard rates or overall AI conception of commercial and pure-
error (0.172 and 0.048 for Exp. 1 and Exp. 3, respec- bred Angus heifers synchronized using a 14-d CIDR-PG
tively) and it is likely this difference is physiologically protocol. Gadberry and Baldridge (2013) also noted no
insignificant. Still, other authors have noted decreased effect on pregnancy rate when Angus cows were treated
performance in heifers treated with an injectable trace with either an injectable trace mineral or no injection.
mineral after stress due to shipping or transportation Although the authors speculated the greatest response
(Arthington et al., 2014). Arthington et al. (2014) hy- would be noted in this experiment, as heifers were less
pothesized this performance decrease is due to an in- than the recommended 60% of mature BW, control heif-
crease in Cu-dependent acute phase protein ceruloplas- er still had acceptable AI pregnancy rates (66%). Other
min, which can affect nutrient metabolism and animal researchers have reported favorable reproductive suc-
growth (Johnson, 1997), and could be greater in cattle cess when heifers were bred at 50 to 55% of mature BW
with an increased Cu status. From a management stand- (Funston et al., 2012; Gunn et al., 2015).
point, it is unlikely the heifers in the current experi- Exp. 2 heifers treated with a trace mineral injection
ments, had been subjected to any stress that would have (TMI) tended to have greater AI pregnancy rates com-
increased these acute phase proteins. However, stress pared with their control counterparts. However, overall
was not assessed in this trial so, it is unknown if this pregnancy rates were similar regardless of treatment.
may have explained the BCS changes noted at breeding. Heifers that conceive to AI will not only have an in-
In both Exp. 1 and Exp. 3, pregnancy rates to timed creased probability of weaning more calves in their life-
AI and overall pregnancy rates were similar for both time but also heavier calves (Burris and Priode, 1958).
treatments. In an intensively managed dairy herd fed a Additionally, when more heifers conceive to AI, this can
TMR that met NRC requirements for trace mineral sup- result in a more uniform calf crop and a shortened breed-
plementation, no difference was reported in first-service ing season (Dziuk and Bellows, 1983). Even though all
conception rate when a single injection of trace mineral heifers had access to free choice trace mineral during the
was administered prior to breeding (Vanegas et al., 2004). grazing period and were provided with an organic trace
While it was physiologically unclear why this may have mineral as part of a TMR from weaning until beginning
occurred, this data could suggest additional trace mineral the grazing period, a favorable response (P = 0.07) in
supplementation in a herd provided with NRC recom- AI pregnancy rates due to the trace mineral injection was
mended levels of trace minerals provides no beneficial still noted. Since offering free choice mineral can result
effects. The heifers in the current study were provided in inconsistent consumption and erratic intake, it is pos-
trace mineral at or above NRC recommendations, sug- sible the trace mineral status of MM2 heifers was more
gesting they likely had an adequate mineral status. optimal than CON2 heifers. However, it is important
It is important to note heifers in Exp. 3 weighed 348 to note the distribution of tail paint scores was similar
kg at breeding, approximately 53% of the herd’s 650 across treatments, suggesting all heifers responded simi-
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 Injectable trace mineral heifer reproduction 465

larly to estrus synchronization, and thus conception dif- TMI to improve heifer mineral status at breeding, when
ferences were not due to differences in synchronization trace minerals are of utmost importance. This could
response. Interestingly, Kirchhoff (2015) reported a simi- help to explain why increased AI conception rates were
lar increase in pregnancy rate to AI when beef heifers noted for MM2 heifers over their CON2 counterparts.
were treated with an injectable trace mineral and noted Unfortunately, since trace mineral status was not as-
no differences across treatment in estrous behavior as sessed in this trial, we are unable to conclude if these
indicated by estrous detection patches. Mundell et al. Simmental heifers did have a greater need for the trace
(2012) also noted comparable results when heifers were minerals provided from the trace mineral injection.
treated with either a trace mineral injection or with sterile Heifers utilized in Exp. 2 were developed in a dry-
saline; heifers that received the trace mineral injection lot and then moved to an early grazing season pasture at
had greater fixed time AI conception rates and overall breeding. The NRC (2016) suggests this management
pregnancy rates were similar between treatments. These strategy can result in BW loss and greater instances of
data suggest an injectable trace mineral may increase AI reproductive failure. The heifers in Exp. 2 lost 13.5 kg

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conception rates even if dietary trace minerals are meet- from the initiation of the trial to breeding (33 d), sug-
ing heifer’s requirements. gesting the negative energy balance immediately prior
However, as with performance parameters, the ef- to breeding could have decreased reproductive perfor-
fects of an injectable trace mineral on reproductive per- mance. It is possible that the additional Cu, Mn, Se, and
formance has been inconsistent. Brasche et al. (2014a) Zn provided to the MM2 treated heifers could have
treated Angus heifers with either a trace mineral or ster- played an important role in maintaining reproductive
ile saline injection 33 d prior to AI and reported greater success. Trace minerals play a crucial role in a ruminant
AI and overall pregnancy rates for heifers receiving a animal’s productivity (Suttle, 2010) and can play im-
trace mineral injection. These differences were noted portant roles in hormone synthesis (Paterson and Engle,
despite the trace mineral injection having no effect 2005), which is vital for reproductive success. However,
on the liver concentration of Mn, Cu, or Zn. Though, further research is needed to elucidate how trace miner-
Brasche et al. (2014a) did report an increase in liver Se als alter or improve embryo survival when cattle are in
concentrations of heifers treated with a trace mineral a negative energy balance at breeding.
injection. In contrast, Daugherty et al. (2002) improved In conclusion, under the conditions of these 3 ex-
the Cu status of cows treated with trace mineral injec- periments, an injectable trace mineral administered 33
tion and vitamin E; however, saw no effect on the con- d prior to the breeding season, in certain management
ception rate of beef cows compared to saline-treated settings, resulted in increased AI conception rates of
cows. Brasche et al. (2014b) also noted no difference heifers even when provided adequate trace mineral sup-
in AI or overall pregnancy rates when cows were given plement. Due to the difficulty of assessing trace mineral
either a trace mineral or saline injection at breeding. status of an entire herd, supplementing trace minerals
While it is unknown what may be driving this variabil- through an injection may be a viable way to ensure a
ity across experiments it is possible other factors such consistent, adequate trace mineral supply to heifers
as breed differences, management strategies, nutrient for optimal reproductive performance. The reproduc-
status, and potential mineral antagonists are playing a tive response across these 3 experiments was variable.
complex role in assessing reproductive performance in However, it is important to note that injectable trace
cattle supplemented with an injectable trace mineral. minerals do not appear to incur any negative impacts on
The cattle utilized in Exp. 2 were predominantly heifer performance or reproductive success. Further re-
Simmental influenced cattle, while those utilized in search is required to substantiate this hypothesis and to
Exp. 1 were Angus, suggesting breed differences may further understand the response differences elicited by
account for some variability noted. Particularly with Cu, injectable trace mineral supplementation across herds.
differences have been reported in how Simmental cattle
absorb (Ward et al., 1995) and excrete (Gooneratne et LITERATURE CITED
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