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Animal Reproduction Science 86 (2005) 305–316
Plasma progesterone concentration dependson sampling site in pigs
Juha V. Virolainen
a
,
b
,
∗
, Robert J. Love
b
, Anssi Tast
a
,Olli A.T. Peltoniemi
a
a
 Department of Clinical Veterinary Sciences, Faculty of Veterinary Medicine,University of Helsinki, Pohjoinen Pikatie 800, 04920 Saarentaus, Finland 
b
 Department of Clinical Veterinary Sciences, Faculty of Veterinary Science,University of Sydney, Camden, NSW 2570, Australia
Received 11 March 2004; received in revised form 13 July 2004; accepted 20 July 2004
Abstract
The objective of this study was to examine a possible difference in progesterone concentrationsbetweenthesystemicvenousbloodandthecaudalvenacavainearlypregnantgilts.Nineteencrossbredpregnant gilts were offered three different regimens of feeding to examine influence of feeding onthe secretion pattern of progesterone. The groups were high (H–H), low (L–L) and low–high (L–H)receiving 3.6, 1.8 and 1.8/3.6kg/day, respectively. Catheters were placed in a jugular vein and thecaudal vena cava (to sample ovarian secretion) on day 19 of pregnancy. Two consecutive samplestaken at 30-min intervals were collected four times a day for 5 days (days 20–24). In addition,three gilts were simultaneously sampled from both catheters at 30-min intervals for 12h on day 22.Progesterone concentration was significantly lower in the jugular vein compared with the caudalvena cava in all three feeding groups (
P
< 0.001). An indication of episodic pattern of progesteroneproduction occurred in plasma collected from the caudal vena cava, but not from the jugular vein.Dietary intake did not cause a profound effect on plasma progesterone concentrations during days20–24 of gestation. It seemed that ovarian progesterone was released into the vena cava in an episodicpattern and there were implications that these episodes were temporally associated with LH pulses.© 2004 Elsevier B.V. All rights reserved.
Keywords:
Luteinizing hormone; Mechanisms of hormone action; Ovary; Pregnancy; Progesterone
∗
Corresponding author. Tel.: +358 19 5295323; fax: +358 19 6851181.
 E-mail address:
juha.v.virolainen@helsinki.fi (J.V. Virolainen).0378-4320/$ – see front matter © 2004 Elsevier B.V. All rights reserved.doi:10.1016/j.anireprosci.2004.07.004
 
306
J.V. Virolainen et al. / Animal Reproduction Science 86 (2005) 305–316 
1. Introduction
Progesterone is considered to orchestrate uterine secretions, which in turn arecrucial for embryonic development and growth (Adams et al., 1981; Vallet et al., 1998).Low feed intake in the immediate postmating period is associated with elevated pro-gesterone concentrations and with improved embryonic survival (Jindal et al., 1996).Progesterone levels may be subjected to seasonal variation (Wrathall et al., 1986), pos-sibly due to seasonally suppressed secretions of luteinizing hormone (LH). These hor-monal changes during the late summer and early autumn might result in loss of thewhole litter, which is known as one of the seasonal infertility manifestations (Love,1978). In contrast to findings byJindal et al. (1996), high feeding rate immediately after mating has been reported to provide benefits for reproduction.Love et al. (1995)reported that liberal feeding improved pregnancy rate during the known seasonal in-fertility period. Although those benefits are considered to be mediated by LH andprogesterone, it appears that progesterone concentrations in the peripheral blood are neg-atively influenced by a high feed intake, also at the time of seasonal infertility, eventhough increased LH secretion was detected at the same time (Virolainen et al., 2004).In spite of the decreased progesterone level, the pregnancy rate was significantly higherin gilts with abundant feeding as compared with the other regimen (Virolainen et al.,2004).Progesterone concentration in the circulating blood reflects the balance between syn-thesis and metabolic clearance of progesterone. A high food intake results in increasedportal blood flow and metabolic clearance of progesterone and is, therefore, linked tolower progesterone concentration in plasma of sheep (Parr et al., 1993)and pigs (Prime and Symonds, 1993). However, it is possible that the production of progesterone in-creases due to a high rate feeding, but the rise is negated by an increased portal bloodflow and metabolism. The benefits of increased progesterone production could be medi-ated locally to the uterus by countercurrent transfer of progesterone from venous bloodfrom the ovary into arterial blood supplying the uterus as demonstrated in early pregnantsows (Stefanczyk-Krzymowska et al., 1998).Krzymowski et al. (1986)demonstrated a close anatomical association and specific structures between the venous, arterial and lym-phatic vessels along the mesometrium, which might enable penetration of steroids into theuterine arterial blood. The described mechanism might ensure an increased progesteroneconcentration in blood flow to the uterus. High food intake might improve progesteroneproduction, which in turn might have a local effect on the uterus, since a decrease in pro-gesterone concentration of the peripheral blood has been reported (Virolainen et al., 2000,2004).However, catheterization of the utero-ovarian vein requires an intensive surgical proce-dure that might affect ovarian and uterine function. Therefore, catheterization of the caudalvena cava via the lateral saphenous vein would be an appropriate method to monitor pro-gesterone production.The objective of this study was to examine the difference in progesterone concentrationsin the jugular and vena cava blood in early pregnant gilts. Furthermore, three feedinglevels were used to determine if nutrient intake had an effect on ovarian progesteroneproduction.
 
 J.V. Virolainen et al. / Animal Reproduction Science 86 (2005) 305–316 
307
2. Materials and methods
2.1. Animals, management and feeding
Nineteen crossbred (Landrace and Large White) pregnant gilts (138
±
15.7kg, mean
±
S.D.) from a commercial farm (Hillcrest Park piggery, Menangle, NSW, Australia) wereused in this study. Gilts were transferred to light (12h light and 12h dark) and temperature(+23
±
1
◦
C) controlled climate rooms in two groups (8 and 11 gilts) with 6-week interval,5–9 days after natural mating (day 0). Gilts were weighed after arrival and at the end of the trial. The gilts were housed in individual stalls and fed once a day at 9.00 a.m. witha commercial diet (Amitie Dry Sow diet; Vella Stock Feeds, Plumpton, NSW, Australia;13MJ/kg)of2.5kg/dayfromarrivaluntilday12or13ofpregnancy.Theanimalswerethenrandomlyassignedtothreeregimens,whichwerechosenbasedonearlierstudiesofauthors(Peltoniemi et al., 1997a,b;Virolainen et al., 2004). From day 12 or 13, high feeding group (H–H) received 3.6kg/day until the end of trial (day 24), low feeding group (L–L) received1.8kg/day and the low–high feeding group (L–H) had low rate (1.8kg/day) until day 21of pregnancy followed by a high rate feeding (3.6kg/day) thereafter. Low level was aboutequal to maintenance requirements (metabolic body weight
∗
0.458MJ) and high level was2.5 times maintenance per day. All pigs were tested for pregnancy by real time ultrasound.The sampling period, 20–24 days after mating, was chosen to minimize daily variations inprogesteroneconcentration,sinceprogesteroneconcentrationappearstoremainreasonablyconstant beyond day 20 (Tast et al., 2000).
2.2. Blood collection
Gilts were fitted nonsurgically with an indwelling jugular vein cannulae via an ear veinon day 19 of pregnancy (Peacock, 1991).Gilts were fastened around the upper jaw with a rope snare and tied to fence while an i.v. catheter placement unit (14G, 57mm) wasinserted into an ear vein and a vinyl tube (o.d. 1.5mm
×
i.d. 1.0mm) was passed throughthe catheter. The free tube was secured to the ear with adhesive tape wrapped around theear and the ear was fixed to the neck by a collar of adhesive tape. The free end of tubewas stored in a Velcro pouch attached to the neck for an easy sampling. The time taken forcatheterization was approximately 5–10min.In preparation for catheterization of the caudal vena cava (Benoit and Dailey, 1991),anesthesia was induced with an i.v. bolus injection of thiopentone sodium at a dosage of 1.25g diluted in 25-ml sterile water. Anesthesia was maintained with halothane/oxygenmixture delivered by a facemask. Gilts were laid on the left side and lateral aspect of theright rear leg shaved approximately 20cm dorsal to the hock and the area was disinfectedwith 70% EtOH. An incision was made through the skin approximately 3cm dorsal to thehock and 2cm lateral to the Achilles tendon. The subcutaneous fat (1–2cm) was separatedand the lateral saphenous vein (SV) isolated by blunt dissection. Tweezers were placedunder the vessel and two loose ligatures were inserted approximately 1cm apart. Slighttraction was maintained on distal ligature, a small transversal incision made in the veinand a catheter (1.0mm i.d., 1.5mm o.d.) was inserted. Each catheter was marked previ-ously at 4-cm intervals from 48 to 56cm from the tip. Plasma samples were collected at
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