Technical note: A new surgical technique for

ovariohysterectomy during early pregnancy in beef heifers1

K. J. McLean,*2 C. R. Dahlen,* P. P. Borowicz,* L. P. Reynolds,*
M. R. Crosswhite,* B. W. Neville,† S. D. Walden,‡ and J. S. Caton*
*Department of Animal Sciences and Center for Nutrition and Pregnancy, North Dakota State University,
Fargo 58102; †Central Grasslands Research and Extension Center, North Dakota State University, Streeter 58483;
and ‡Office of the Vice President and University Attending Veterinarian, North Dakota State University, Fargo 58102

ABSTRACT: We hypothesized that a standing flank blind palpation and ligate them through the broad liga-
ovariohysterectomy procedure could be developed in ment via an improved clinch knot, 3) cut the ovaries
beef heifers that would provide high quality tissues for and uterus free from the broad ligament, 4) ligate the
addressing critical questions during early pregnancy, cervix and uterine branch of the vaginal artery, and 5)
while concomitantly keeping livestock stewardship a cut through the cervix and remove the reproductive
high priority. To test the hypothesis, we: 1) developed tract. Surgical times, from skin incision to place-
a standing flank ovariohysterectomy procedure for use ment of the last suture, were influenced (P = 0.04) by
in beef heifers, and 2) implemented this procedure in stage of gestation. In pregnant heifers, time decreased
a cohort of heifers up to d 50 of pregnancy for tissue from d 22 (120.0 ± 12.0 min) of gestation to d 40
collections, documentation of post-surgical recov- (79.5 ± 12.0 min) of gestation; then increased at d 50
ery, and assessment of feedlot finishing performance. (90.5 ± 14.7 min) of gestation. Using this procedure,
Ovariectomy and cesarean section protocols are well we obtained uterine, placental, and embryo/fetal tis-
established in research and veterinary medicine and sues that had experienced limited hypoxia, little or no
were used as starting points for procedural develop- trauma, and thus were excellent quality for scientific
ment. Crossbred Angus heifers (n = 46; ~ 15 mo of study. All heifers recovered from surgery quickly and
age; BW = 362.3 ± 34.7 kg) were used to develop this were moved to a finishing period. During the finish-
new surgical tissue collection technique. Heifers were ing period, ovariohysterectomized heifers had a DMI
subjected to the 5-d CO-Synch + CIDR estrous syn- of 13.8 kg, gained 1.99 ± 0.35 kg/d, and had a G:F
chronization protocol so ovariohysterectomy occurred of 0.145 over 132-d. The standing flank ovariohyster-
at d 16, 22, 28, 34, 40, and 50 of gestation. Key aspects ectomy technique represents a new and viable model
of the standing flank ovariohysterectomy technique to economically obtain high quality tissues for inves-
included 1) use of local anesthetic for a standing flank tigating critical biological mechanisms during early
incision, 2) locate the uterine and ovarian arteries via pregnancy in beef heifers.
Key words: bovine, early gestation, ovariohysterectomy

© 2016 American Society of Animal Science. All rights reserved. J. Anim. Sci. 2016.94:5089–5096
doi:10.2527/jas2016-0761
INTRODUCTION
1Appreciation is expressed to personnel at the NDSU Animal

Nutrition and Physiology Center, NDSU Animal Science Nutrition The world needs to increase its output of meat
Laboratory and NDSU Animal Science Reproductive Physiology protein by 2050 on the same resources to meet the
Laboratory for their assistance in completing this project. A special projected human food demands (Reynolds et al.,
thanks to Arlyn Scherbenske for providing additional veterinary 2015); thus, efficient livestock production is essential.
medical consultation during the initial phase of procedural develop- Embryonic development depends on healthy placen-
ment, Faithe Keomanivong for drawing all schematics depicting tal formation but currently there are no data showing
harness and ligature placement, and Jamie Keomanivong for how placental insufficiency occurs in cattle. Impaired
computerizing all harness and ligature schematics. pregnancies are associated with early embryonic loss-
2Corresponding author: kyle.j.mclean@ndsu.edu
es (Reynolds et al., 2014) and long-term effects on the
Received June 28, 2016.
offspring (Reynolds and Caton, 2012).
Accepted October 5, 2016.
5089
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 5090 McLean et al.

Bovine placental tissues are collected by embryo (1.38 g of progesterone; Zoetis), followed in 5 d by
flushing up to d 16 (King et al., 1981). Due to damage CIDR removal and 5 mL of prostaglandin F2α (250 mg
to the uterus and embryo size restrictions, slaughter ap- dinoprost tromethamine [Lutalyse] i.m; Zoetis), fol-
proaches are used at d 16 (Cooke et al., 2014) and later lowed in 8 h by an additional 5 mL Lutalyse. Estrotect
in gestation. Slaughter approaches are expensive due patches (Rockway Inc., Spring Valley, WI) were placed
to decreased carcass value because these animals are on the tailhead at the time of CIDR removal to aid in the
not fattened to industry standards. Thus, only limited detection of estrus. Heifers were AI bred to a single sire
information exists for utero-placental development af- 8 to 14 h after estrus was observed (day of breeding = d
ter d 16 of pregnancy in cattle and almost no data exists 0). Ovariohysterectomies were performed on d 16, 22,
between d 16 and 30 of gestation. Most of the 3.67 mil- 28, 34, 40, and 50 (n = 9, 6, 6, 7, 6, and 5 respectively)
lion (USDA-NASS, 2015) heifers destined for feedlots of gestation and at d 16 of the estrous cycle for non-
present an opportunity to study pregnancy establish- bred, non-pregnant controls (NP; n = 7). Pregnancy was
ment and embryonic loss within a normal reproductive confirmed via transrectal ultrasonography on d 22 and
tract with a technique to acquire these tissues. again on the day of surgery (d > 22).
Midventral (Wiltbank and Casida, 1956; Copelin et
al., 1987) or left lumbar (Wiltbank and Casida, 1956; Pre-Surgical Preparation
Hamernik et al., 1985) hysterectomies have been re-
ported in non-pregnant females. However, a procedure To minimize gut fill during surgery, feed and wa-
which allows for collection of pregnancy tissues which ter were withheld for 24 and 12 h, respectively, before
have experienced limited hypoxia and damage during surgery. All items needed for the surgeries are listed in
acquisition would provide a platform to address ques- Table 1. The hair in the surgical field, which was on the
tions regarding embryonic survival and pregnancy out- left paralumbar fossa, was clipped cranially to approxi-
comes. Therefore, we hypothesized that a standing flank mately the 10th rib, caudally 5 cm past the tuber coxae
ovariohysterectomy procedure could be developed in (most lateral, cranial point of the pelvic ilium, some-
bovine heifers that would provide high quality tissues times referred to as the ‘hook’ bone), dorsally to the ver-
for addressing critical questions during early pregnancy, tebral column, and ventrally to femorotibial joint (stifle
while efficiently utilizing animal resources and con- joint) with standard large animal clippers with 83 and
comitantly providing acceptable livestock stewardship. 84AU blades (Oster Shearmaster clippers; Sunbeam
Products; Boca Raton, FL). A number 40 Oster blade
MATERIALS AND METHODS (Sunbeam Products) was used to remove hair down to
the skin cranially to the 11th rib, caudally 2.5-cm past
All animal procedures were conducted with approval the tuber coxae, dorsally 2.5-cm above the transverse
from the Institutional Animal Care and Use Committee processes of the lumbar vertebrae, and ventrally 2.5-cm
at North Dakota State University and under the direct su- below the paralumbar fossa. Heifers were clipped 12
pervision of the university attending veterinarian. h before surgery. On the day of surgery, heifers were
placed into a restraining chute (Silencer Commercial
Animals Pro Model; Moly Manufacturing Inc.; Loraine, KS)
and given 0.05 mg acepromazine maleate (PromAce
Crossbred Angus heifers (n = 46; ~ 15 mo of age; Injectable; Zoetis) per kg BW intravenously.
BW = 362.3 ± 34.7 kg) were transported 229 km from To facilitate animal care and restraint, the heifer was
the Central Grasslands Research and Extension Center then equipped with a cranial (Fig. 1A) and caudal (Fig.
(Streeter, ND) to the Animal Nutrition and Physiology 1B) support harness (lateral view, Fig. 1C) and nylon
Center (North Dakota State University, Fargo, ND). hobbles on the rear legs. These harnesses are vital to
The heifers were housed in group pens with a maximum the efficiency of the technique. The caudal harness (Fig.
of 6 heifers per pen. The goal of the experiment was to 1B) consisted of two continuous loop 1.8 m tow straps
develop an ovariohysterectomy technique to facilitate (Roundsling; Lift-all; Landisville, PA). The left side tow
high quality tissue collections for use in studying bio- strap began dorsal to the sacral vertebrae, progressed ven-
logical mechanisms and events occurring during early trally down the femur and caudal to the tuber coxae, along
pregnancy. Before surgery all heifers were subjected to the medial side of the tibia and dorsal again caudal of the
the 5-d CO-Synch + CIDR estrus synchronization pro- femur to rejoin the other end of the strap. The right side
tocol (Bridges et al., 2008). Heifers received 2 mL of tow strap also began dorsal to the sacral vertebrae, pro-
gonadotropin releasing hormone (100 µg gonadorelin gressed ventrally down the femur and cranial to the tuber
hydrochloride [Factrel] i.m.; Zoetis; Florham Park, NJ) coxae, along the medial side of the tibia and dorsal again
and a controlled internal drug releasing (CIDR) insert caudal of the femur to rejoin the other end of the strap. The

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Table 1. List of materials needed for the ovariohyster-

ectomy procedure
Quantity Item
Surgeon attire and preparation items
1 Sterile scrub top (various sizes)
1 Sterile surgical gloves
1 E-Z scrub hand brushes with providine
Pre-surgical items
1 Standard large animal clipper (83 and 84AU blades)
1 Small clippers with Number 40 blade
1 Bottle of 7.5% povidone-iodine scrub
1 Spray bottle of 7.5% povidone-iodine solution
1 Spray bottle of 70% isopropyl alcohol
36 mL 2% lidocaine hydrochloride local anesthetic
5 mL 2% lidocaine hydrochloride epidural
0.3 mL Acepromazine maleate
1 3 mL syringe
1 6 mL syringe
3 12 mL syringe
1 Sterile chute drape
4 18 gauge 3.8 cm needles
1 20 gauge 3.8 cm needle
1L Benz-All germicidal solution
Surgical kit
2 #4 Scalpel handles
1 #20 Scalpel blades
1 Rat tooth forceps
4 Medium hemostats (18 cm)
4 Mosquito hemostats (12.5 cm
2 Crafoord Coarctation Clamp (23.5 cm)
2 Needle holders Figure 1. Surgical restraint and ovariohysterectomy harness sche-
matic in cranial, caudal and lateral views A) Schematic of the cranial har-
1 Double blunt scissors (28 cm)
ness B) Schematic of the caudal harness and C) Lateral view of incision
2 Double blunt scissors (13 cm) site (red line), cranial harness, and caudal harness (Schematic A, B, and C
1 Single blunt scissors (13 cm) drawn by Faithe Keomanivong and computerized by Jamie Keomanivong).
3 3/8 circle taper needles (#1, #2, or #4)
3 3/8 circle cutting needles (#1, #2, or #4)
each end. The rubber strap was placed ventral to the 6th
1/4 roll 3” × 3” or 4” × 4” gauze pads
2 1/2 circle taper needles with blunted end (#1)
rib and caudal to the fore limbs. The iron triangles were
Surgical items secured via 28-cm cable ties (Lowe’s Company Inc.) and
274 cm #2 Vicryl absorbable suture chained to an iron leg spreader dorsal to the 1st thorasic
1m #1 or #2 heavy polymerized braunamid vertebrae. The leg spreader was attached to a 5-cm ratchet
10 mL 0.01M Phosphate buffered saline strap (Lowe’s Company Inc.) which allowed surgical
1L Sterile saline with 10 mL of glycerol assistants the ability to lift the cranial half of the animal.
anhydrous and 4 mL penicillin The cranial harness was secured forward by 2.5-cm nylon
Post-surgical items straps (Lowe’s Company Inc.) that progressed from the
0.02 mL/kg Benzathine-procaine penicillin G ventral midline cranial to the fore limbs and on each side
0.02 mL/kg Flunixin meglumin (for 3 consecutive d)
of the cervical vertebrae. Straps are twisted 2 or 3 times
around each other dorsal to the cervical vertebrae and
then connected to the leg spreader with straps. The cranial
and caudal harnesses allowed for slight manipulation of
4 ends were connected via a grade-8 U-bolt and a 3 × 10- the animal to maintain an adequate surgical field which
cm steel plate. The U-bolt was attached to a 5-cm ratchet is ideal if the animal is standing. Harnesses also prevent
strap (Lowe’s Company Inc., Mooresville, NC) which al- the animal from lying down which completely obscures
lowed surgical assistants the ability to lift the caudal half the surgical field. This immobility of the heifer allowed
of the animal. The cranial harness (Fig. 1A) consisted of the surgeon to palpate and ligate structures necessary for
a single rubber strap 1-cm thick, 18-cm wide, and 1-m completion of the surgical procedure without major inhi-
long that had an iron triangle 1-cm in diameter bolted into bition from animal movement.

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The surgical field was then scrubbed with 7.5% po- further distal to the uterine horn than could be palpated
vidone-iodine scrub (Betadine; Creative Science LLC; while still in contact with the uterine horn and no more
Ballwin, MO) and rinsed with 70% isopropyl alcohol proximal to the uterine horn than the 1st arterial branch
(Vetone; Boise, ID). For local anesthetic block 36 to from the main uterine artery. In addition, care was taken
42 mL of 2% lidocaine hydrochloride (approximately 4 so that any branches of the uterine artery were included
mg/kg BW; Vetone) was administered with an 18 gauge within the ligature to ensure complete blockage of blood
3.8-cm-long needle (Medtronic Inc., Dublin, Ireland) flow. This process was repeated for the right uterine ar-
subcutaneously and into the intercostal muscles in an in- tery, both ovarian arteries, and the cervix. For the cervix,
verted L-pattern starting 5 cm cranial to the tuber coxae care was taken so that the uterine branch of the vaginal
and continuing cranially along the transverse processes artery on both sides was included within the ligature.
of the lumbar vertebrae to the 13th rib, then ventrally Separation of the broad ligament was initiated by
along the 13th rib to the ventral portion of the para- creating an opening large enough for the surgeon to
lumbar fossa (Turner and McIlwraith, 1982), and then insert 2 fingers caudal to all ligatures. Once the initial
as needed to maintain adequate local anesthesia. After opening was created surgeons were able to completely
administration of the local anesthetic, the surgical site localize and protect cutting target tissue in their hand.
was scrubbed with 7.5% povidone-iodine scrub, rinsed The broad ligament was then completely severed from
with water, and coated with ethanol and 7.5% povidone- the ovaries, uterine horns, and uterine body to include
iodine solution and allowed to dry. Anesthetic block (Ko no less than 2 cm of the cervix. Severing of the broad
et al., 1989) to the uterine body, uterine horn, and broad ligament with the double blunt scissors was conducted
ligament was administered just before the initiation of in a manner so that all other internal organs and tissues
surgery. The block was completed by injection of 5 mL were protected from the cutting surfaces. As gestation
of 2% lidocaine hydrochloride with an 18 gauge, 3.8-cm- progressed and blood flow increased, a 2nd ligature
long needle (Medtronic Inc.) in the epidural space of the was placed distal to the first ligatures on all 4 uterine
tail head between the second and third sacral vertebrae. and ovarian arteries. This order of artery ligation and
broad ligament separation seemed to minimize tissue
Surgical Procedure edema during completion of surgery by decreasing
overall time for removal of tissues. Minimizing ede-
Instruments included in the sterilized surgical kits ma is important because edema presence decreased
and materials for surgery are listed in Table 1. After pre- the ability to palpate the uterine, ovarian, and cervical
surgical preparation, an incision, approximately 18 to 25 structures. An improved clinch knot was placed on the
cm in length (i.e., wide enough to accommodate the 2 cranial end of the cervix (Fig. 2A) and a second im-
arms of the surgeon side-by-side) was made in the left proved clinch knot was placed on the caudal end of the
paralumbar fossa. The incision began 3 to 5 cm ventral uterine body (~ 1 cm apart).
of the transverse processes of the lumbar vertebrae and After placement of cervical ligatures, the uterine
3 to 5 cm cranial to the tuber coxae and progressed ven- body was clamped via a 24-cm Crafoord Coarctation
trally at an approximate 25° angle cranially and end 3 Clamp (Integra-Miltex) which was placed (Fig. 2A)
to 5 cm caudal to the 13th rib in the ventral region of cranial to the cervical ligation(s). Clamp placement
the paralumbar fossa. The external and internal abdomi- was as close as possible to the cervical ligation(s),
nal oblique, transverse abdominal muscles, and perito- while still allowing for adequate space to complete
neum were then incised or blunt dissected to gain ac- the incision, and across the entire uterine body. Clamp
cess to the peritoneal cavity. Once inside the peritoneal placement ensured that fetal membranes remained in
cavity the rumen, small intestine, rectum, reproductive place with no damage on removal of the reproduc-
tract, and broad ligament were palpated to confirm nor- tion tract. At this stage of gestation, fetal membranes
mal anatomical structures and locations. The left uter- were small enough to avoid damage on removal but
ine artery (closest to incision) was located and ligated as gestation continues damage to chorion at the cra-
(Fig. 2A) with an improved clinch knot (Fig. 2B: http:// nial cervical os may occur. An incision was made be-
hdimagegallery.biz/improved+clinch+knot+fishing) us- tween the uterine clamp and the 2nd cranial cervical
ing #2 absorbable suture (Vicryl; Ethicon; Somerville, ligature to separate the uterine body from the cervix
NJ). Suture material was passed through the broad liga- (Fig. 2A). This incision was made using the 28-cm
ment caudal to the uterine artery back through broad double blunt scissors (Integra-Miltex) and using the
ligament cranial to the uterine artery, with either (sur- uterine clamp as a guide. The target tissues (ovaries
geon preference) a blunted #1, half curve, taper needle and uterus) were then removed from the peritoneal
(Integra-Miltex; Plainsboro, NJ) or 13-cm hemostats cavity, placed on a tray, and covered with cheesecloth
(Integra-Miltex). The ligature was placed (Fig. 2A) no moistened with 0.01M PBS. Upon removal of the ova-

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 Bovine ovariohysterectomy 5093

Figure 2. (A) Schematic of bovine reproductive tract illustrating the location of ligatures (black line: atertial ligatures, green line: cervical ligatures), clamp
(gold line: 24-cm Crafoord Coarctation Clamp; Integra Miltex), and uterine incision (dashed line) during the ovariohysterectomy procedure and (B) Close up
schematic of the improved clinch knot around the ovarian artery (knot adapted from http://hdimagegallery.biz/improved+clinch+knot+fishing; schematic A and
B drawn by Faithe Keomanivong and computerized by Jamie Keomanivong).

ries and uterus, the rumen and intestines were palpated individual recovery pens and provided grass hay and
to ensure no damage had occurred, and 1 L of sterile water, ad libitum. Antibiotic (10,000 units of benzathine-
saline (~38.6° C) containing 10 mL of glycerol anhy- procaine penicillin G/kg BW; PenOne Pro; Vetone) was
drous (J. T. Baker Chemical Co.; Avantor Performance given for 2 consecutive d following surgery. Heifers re-
Materials; Center Valley, PA) and benzathine-pro- mained in their recovery pen for no less than 5 d and no
caine penicillin G (10,000 units/kg BW; PenOne Pro; more than 10 d, after which they were returned to group
Vetone) was administered into the peritoneal cavity. pens. Sutures were removed from the skin 14 d follow-
The peritoneum and abdominal muscles (transverse ing the surgery. After recovery and project completion,
and oblique) were closed with #2 absorbable suture all heifers were placed in a feedlot on a single d and fed
(Vicryl; Ethicon) via a continuous Ford interlocking a high-concentrate diet, which consisted of 50% course
pattern (Knecht et al., 1987). The skin was closed with rolled corn, 25% dried distillers grains with solubles,
a #2 polymerized braunamid (Jorgensen Laboratories; 20% corn silage, and 5% trace mineral premix, all on
Loveland, CO) via interrupted sutures to ensure inci- a DM basis. The heifers were given an electronic id tag
sion remained closed even after edema. Incision site and fed using an Insentec roughage intake control sys-
and sutures were dressed with Blu-Kote antibiotic tem (Hokofarm Group, Marknesse, Netherlands) until
wound dressing (H. W. Naylor Co., Morris, NY). they reached slaughter weight (~591 kg). Intake was
recorded daily, and BW was taken every 28 d so that
Post-Operative Care ADG and efficiency could be determined.

Antibiotics (10,000 units of benzathine-procaine Procedure Assessment

penicillin G/kg BW; PenOne Pro; Vetone) and anal-
gesics (1.1 mg of flunixin meglumin/kg BW; Prevail The ovariohysterectomy technique was assessed by
Flunixin; Vetone) were administered immediately fol- statistical analysis of length of surgery, stage of gestation,
lowing surgery. Heifers were then placed in 3 × 3-m, and experience of the surgeon. There were 3 primary sur-

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 5094 McLean et al.

geons with multiple trainee assistants with varying de- Dakota State University, Fargo, personal communi-
grees of expertise. However, there was no difference in cation). Until the development of our standing flank
surgery times between surgeons (P > 0.05); thus, length ovariohysterectomy technique, the only way to ob-
of surgery was recorded and reported as means ± SEM. serve interactions between the developing conceptus
Observations of adverse side effects and prolonged and maternal tissues, after d 16 of gestation, was to
recovery were also recorded. All statistical analyses slaughter the dam and excise the entire reproductive
were conducted using SAS (SAS Inst. Inc., Cary, NC). tract. However, as described by Cooke et al. (2014)
Differences between wk of surgery or d of gestation and and Bilby et al. (2004), due to the time required to
time to completion were evaluated via PROC GLM with obtain the reproductive tract after exsanguinating the
significance at P < 0.05. The relationship between wk dam, the fetus and reproductive tissues are likely hy-
of surgery and time to completion was evaluated using poxic. The time to procure sample tissues has been re-
PROC CORR. The success of the surgical procedure was ported to be as little as 15 min post-slaughter (Riding
also evaluated by the presence and length of adverse side et al., 2008), with tissue preparation and collection
effects in the ovariohysterectomized heifers, which were occurring thereafter. In the average commercial abat-
recorded but not statistically analyzed because of the ex- toir the time from exsanguination to the time at which
ceptionally low incidence. the reproductive tracts are removed and sampled can
be longer than 30 min (K. R. Maddock-Carlin, North
RESULTS AND DISCUSSION Dakota State University, Fargo, personal communica-
tion). Espina and Mueller (2011) reported that preana-
Length of surgery decreased as the surgeon be- lytical variability due to tissue damage from oxidative,
came more familiar with the protocol. There was a hypoxic, and metabolic stress, which can occur within
35-min decrease between the first 21 heifers (121.5 ± 20 min of blood supply removal, was a major contribu-
6.7 min) compared with the last 21 heifers (86.6 ± 6.7 tor to variability in molecular research outcomes. The
min) that were ovariohysterectomized. The progres- tissue samples obtained via the current technique were
sive decrease in length of surgery was also evident as removed and sampled within minutes after complete
a decrease in the time needed to complete the surgery ligation of the entire blood supply (i.e., cervical liga-
by wk. The time to complete surgery by wk tended tion). The reduced time from cessation of blood flow
to differ (P = 0.09) among weeks, decreasing from to obtaining the reproductive tract for sampling using
136.3 ± 17.3 min on wk 1 to 79.0 ± 13.4 min on wk 9. the current procedure is therefore a distinct advantage
There was also an inverse correlation between time to for assessing biological events and mechanisms asso-
completion and wk (r = -0.53; P = 0.0009). The length ciated with early pregnancy outcomes.
of surgery was also influenced by day of gestation (P = Before development of our ovariohysterectomy
0.04). Time was least in non-bred, non-pregnant con- technique, the most common way to acquire fetal
trols (69.0 ± 20.8 min). In pregnant heifers, time de- tissues before implantation was by embryo flush-
creased from d 22 (120.0 ± 12.0 min) to d 40 (79.5 ± ing. Techniques to flush developing embryos from
12.0 min). However, the times for d 16 and d 28 may uteri were established in the late 1970s (Testart et al.,
be skewed, as the first surgeries were conducted on d 1975). However, successful embryo flushes cannot be
28, which raised the overall average for that d, and ap- accomplished after about d 16 of gestation (Winters
proximately half of the d 16 surgeries were performed et al., 1942; King et al., 1981) due to tissue damage
at the end of the study. which make laboratory analyses and growth assess-
The time of surgery was influenced by d of gesta- ments difficult if not impossible. Embryos flushed past
tion and likely by effective animal restraint. As most of d 16 are likely damaged because the embryo can oc-
this technique was conducted outside of the surgeons’ cupy space in both uterine horns at this time of gesta-
visual field, tissue edema and associated inflammation tion (Winters et al., 1942; King et al., 1981), are too
were a concern because they made palpation of target large to fit through a foley catheter, and are sensitive to
structures more difficult, which, in turn, increased the damage from uterine manipulation. The present ovar-
time needed to complete the ovariohysterectomy. With iohysterectomy technique also allowed us to obtain
the development of adjustable belt-type harnesses for both maternal as well as intact fetal tissues after d 16.
front and rear legs, effective animal restraint allowed For procurement of tissues after d 16, either slaugh-
the surgeons to more efficiently complete the proce- ter or midventral surgical techniques have been used.
dure and likely decreased the length of the surgery. Issues associate with slaughter techniques were dis-
Tissues acquired by this technique were of equal cussed above. The major repercussion to a midventral
or greater quality to those obtained via slaughter pro- surgery is the need for general anesthesia. However, the
cedures (L. P. Reynolds and P. P. Borowicz, North current technique can be accomplished via local anes-

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thesia of the paralumbar fossa and caudal epidural. This tle working facilities with little modification. Tissues
eliminates issues associated with general anesthesia in acquired using the standing ovariohysterectomy pro-
ruminants such as dealing with salivation and frequent cedure we have developed are of high quality and ex-
regurgitation as well as preventing hypoventilation due hibit minimal trauma. The quality of these tissues has
to excess gas and lung compression (Dunlop and Hoyt allowed our group to characterize expression patterns
Jr., 1997.). The use of local anesthesia also minimizes of endogenous retroviruses (McLean et al., 2016a;
equipment needs by removing the need for ventilators, b), nutrient transporters (Crouse et al., 2015; 2016b),
intubation equipment, oxygen supply, and fluid therapy redefine the distribution of GLUT 3 in reproductive
during surgery. The current technique also minimizes tissues (Osei et al., 2016), determine the existence of
postoperative care which is more extensive after a GLUT 14 in the maternal tissues (Crouse et al., 2016a),
general anesthesia surgery including but not limited to and assess the influences of nutritional restriction on
regaining consciousness (mainly dealing with dorsal endogenous retroviruses and nutrient transporters
and lateral recumbency) and resumption of eating and (Crouse et al., 2016b; c; McLean et al., 2016b). The
drinking (i.e., gastrointestinal function). standing ovariohysterectomy procedure thus provides
All heifers recovered from the surgical proce- an excellent method to conduct early pregnancy re-
dures exceptionally well. Heifers were considered re- search while maintaining livestock stewardship. High
covered when they presented with no elevated body quality tissues can be acquired with minimum cost,
temperature, resumed previous DMI and water intake, and heifers undergoing the ovariohysterectomy can be
and moved without discomfort. Most heifers were eat- expected to reach slaughter BW with minimal effects
ing and drinking within 2 h after surgery, with the few on growth performance. The data acquired using the
exceptions taking no more than 8 h. Sutures were re- present ovariohysterectomy procedure will therefore
moved from all heifers 14 d post-surgery. Of the 46 minimize inefficiencies. This procedure represents an
heifers that underwent ovariohysterectomies, only two excellent model for studying critical events during
(4.8%) exhibited stiffness in the front limbs due to the early gestation in heifers. This model will provide in-
harness, which dissipated after 1 to 2 d. One heifer sight into the effects of maternal nutrition, impacts of
(2.4%) showed signs of vaginal infection 4 wk after individual nutrients, nutrient transporters, the role of
surgery; however, with antibiotic treatment no further endogenous retroviral elements, and lead to the eluci-
incidence was seen. There was a 0% death loss as- dation of the underlying mechanisms associated with
sociated with this cohort of heifers during technique the establishment and maintenance of pregnancy in
development and all heifers resumed BW growth that beef heifers.
was occurring prior to surgery. The lack of prolonged
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