Les Griel called the meeting to order at 8:20 am, and suggested that we deviate slightly from the written agenda and allow remarks from our Administrative Advisor, Dr. Berndtson, and from the USDA/CREES representative, Dr. Hamernik, first to permit time for the students from WVa to arrive. There was mutual agreement for this change.

Bill Berndtson indicated that NERA had approved the new 5-year proposal submitted by our group, but that he was awaiting word from the USDA following a second external review. In light of the title change to the proposal, Bill also indicated that our regional project designation would change from NE-161 to NE-1007. He also indicated that the minutes of our present meeting were due within 60 days. The minutes should be submitted electronically using a template that Bill will provide to us. Bill also emphasized the importance of impact statements in future correspondence from the regional project.

A question was raised concerning whether or not Bob Duby (UMass) planned to continue his participation with the regional project. Discussion concerned speculation of Bob Duby retiring even though he had submitted information for the new 5-year proposal.

Joy Pate expressed her appreciation to Bill B. for compiling the submission of our regional proposal in web-template form and working closely with the group throughout the renewal process.

The entire group thanked Joy Pate for all of her hard work in bringing the pieces of the proposal together, writing and revising, and completing these tasks in such an expeditious fashion. Keith Inskeep commented that the project would not have received approval without Joy’s efforts.

The group welcomed Dr. Debora Hamernik as the new USDA/CSREES representative to our regional project.

Deb provided an update on recent changes in the USDA/CSREES and insight concerning budgetary issues over the next 2 years (Yellow Handout from USDA). Among the changes, there are three new personnel: Dr. Joseph Jen, Dr. Gary Cunningham, and Dr. Peter Burfening. Dr. Jen is the Under Secretary of Agriculture for Research, Education, and Economics. Deb indicated that Dr. Jen plans to place additional emphasis on agricultural genomics in the next few years and is working toward increasing the budget for NRI funding. Dr. Cunningham is the Associate Administrator for CSREES. Dr. Burfening is the Program Director for IFAFS and for Animal Health (Bacteriology & Aquaculture) in the NRI.

Regarding the CSREES budget, Deb indicated that the President’s proposed request for NRI funding in FY2003 is double that of FY2002 ($240 M compared to $120 M). However, other budgets (e.g., Special Grants, IFAFS, etc.) have been reduced considerably or eliminated entirely. Joanne F. questioned how likely the appropriations would be modified in Congress. Deb indicated that it remains to be determined whether, following debate and revision in the legislature, the NRI budget will remain intact. Deb also pointed out that budgetary appropriations to Homeland Security remain strong and should be considered as a potential source of future funding.

Deb also notified the group of the upcoming changes concerning deadlines for submitting applications. For FY2003, section 41.0 (Animal Reproduction) has a deadline of December 1, 2002. Applications must be received by this deadline, as opposed to postmarked by this deadline (as in previous years). She also recommended FedEx overnight delivery rather than US mail because of the ‘anthrax fiasco’ that occurred with mailed-in applications last year.

WVa students and other WVa attendees arrived at this time and were introduced.

Station reports related to all objectives of old project.

Paul Tsang and Dave Townson reported that the manuscript under review during the last NE-161 meeting has been published (JAS 2002 80:1053-1058). Dave apologized for not having reprints to provide to everyone, but promised to mail copies to each of the stations. He also indicated that the UNH Ag Experiment Station covered the cost of publication.

Ron Butler provided an update on the work concerning the somatotropic axis in transition dairy cows (Project Obj. 2B-Cornell report). In summary, cows that do not ovulate the first dominant follicle postpartum have an increase in NEFA prior to calving that is sustained (refer to figure of Cornell report). In addition, there is an increase in triglyceride accumulation in liver (refer to figure of Cornell report). These effects are hypothesized to result in ovarian resistance to IGF, insulin, and possibly gonadotropins. In vitro studies are proposed to examine the effects of fatty acids in culture medium on estradiol production by pieces of ovarian follicles.

Station Reports not directly related to project objectives

Beth Costine (WVa) reported a study (Item 3A-WVa report) examining early regression of the beef cow corpus luteum as a consequence of decreased metabolism of luteal PGF and increased endothelin-1 (ET-1). Early luteal regression did not involve a reduced ability of the CL to metabolize PGF or activation of the ET-1 system; however, prostaglandin dehydrogenase activity was associated inversely with components of the ET-1 system (i.e., endothelin-1 converting enzyme and endothelin-1 receptor).

Keith Inskeep (WVa) shared new information about the role of luteal PGF in luteolysis (Item 3D-WVa report). The studies entailed placement of intraluteal implants containing indomethacin in ewes to inhibit luteal PGF production. Although serum concentrations of progesterone decreased by day 17 post-estrus in all groups of ewes, the CL of ewes containing indomethacin were significantly heavier than controls. This suggests that a decrease in luteal weight, but not a decrease in progesterone production, requires the production of luteal PGF. An additional experiment was conducted to show that peaks in systemic PGFM following PGF administration, associated with a decrease in CL weight, were not of uterine origin.

Joy Pate (OSU) described a study in which the effects of a GnRH agonist on luteal morphology and function were examined (Item 3C-OSU report). The GnRH agonist increased systemic progesterone production and luteal cross-sectional area when administered on Day 3 of the estrous cycle as compared to the other groups (see line graphs-OSU report). When examined histologically, CL of GnRH agonist-treated cows consisted of large luteal cells with greater diameter, and a tendency toward more large cells than small cells (see Tables-OSU report). This suggests that small luteal cells may become large luteal cells as a result of GnRH administration.

Paul Tsang (UNH) provided information on a recently completed study in which collagenase 3 (MMP-13) was characterized in luteal tissue of cows (Item 3A-UNH report). MMP-13 was found in CL throughout the luteal phase, and was localized to endothelial cells. TNFa stimulated pro-MMP-13 mRNA and protein in luteal-derived endothelial cells in vitro. However, the absence of active MMP-13 as determined by Western blotting suggested that regulators other than TNFa, IFNg and MT1-MMP may be responsible for activation of pro-MMP-13.

Dave Townson (UNH) reported a preliminary study conducted by an undergraduate student in which an attempt was made to manipulate the expression of monocyte chemoattractant protein-1 (MCP-1) in CL in vivo by ablating ovarian follicles (Item 3C-UNH report). The results indicated that while follicular ablation by ultrasound-guided, transvaginal needle aspiration appears to be a feasible technique to eliminate follicles and potentially manipulate systemic estradiol, it did not influence MCP-1 expression or progesterone production by the CL.

An additional report from the WVa group (T.D. Maze) was presented concerning the influence of estradiol on gonadotropin surges in prepubertal beef heifers and the effects of weaning (Item 3C-WVa report). The timing of weaning had no effect on estradiol-induced gonadotropin surges. The responsiveness of the hypothalamus to estradiol positive feedback was established at a relatively early age in these calves and was not affected by short-term weaning. It was concluded that the maturation of the gonadotropin surge center does not limit attainment of puberty in beef heifers.

Lunch Break

Station Reports reports related to objectives of new project

Joanne Fortune and Phil Bridges (Cornell) provided an update on ongoing studies concerning exposure of follicular cells to elevated temperatures (hyperthermic conditions) and its effects on follicular secretion of estradiol, androstenedione, and progesterone (Item B-Cornell report Part II). Exposure of follicle cells to hyperthermic conditions is suggested to affect both basal and gonadotropin-stimulated steroidogenesis. The results thus far suggest that hyperthermia may produce changes similar to premature luteinization and thus might affect fertility in vivo by a similar mechanism.

Ron Butler (Cornell) indicated that their group was beginning an experiment this summer in which they plan to examine seasonal events surrounding follicular development and the follicular to luteal transition that may affect embryonic/fetal mortality (Page 2-Cornell report). Dave Townson commented that some of the methods of the proposed study were strikingly similar to some work just recently completed by 2 undergraduate students at UNH (Item 3B-UNH report). A discussion ensued and Dave agreed to help Ron with Objective 1 of the revised project provided that UNH has cows and some student help available to do this work. Les Griel (PSU) mentioned the possibility of Penn State also participating in this objective.

Ida Holoskova informed the group of a recently completed WVa study in which the effect of immunizing sheep against the peptidoglycan-polysaccharide complex (PG-PS) of Streptococcus pyogenes (a contributing pathogen of mastitis) on pregnancy and mastitis was investigated (Item 3E-WVa report). The presence of IgG to PG-PS in ewe lambs immunized against Streptococcus pyogenes did not diminish early pregnancy loss or the incidence of mastitis. The results were somewhat unexpected, but were consistent with the idea that ewes repeatedly exposed to the pathogen were more likely to have lower pregnancy rates and an increased incidence of mastitis.

Joy Pate suggested that these results were completely consistent with her expectations, explaining that a heightened immune response at the time of insemination would not be expected to result in pregnancy. Further discussion took place and the suggestion was made that this experiment might be repeated, waiting an additional period of time post-immunization before attempting to inseminate the ewes and challenge them immunologically.

Melanie Starbuck presented preliminary data on a study of the effects of growth hormone at insemination on conception rates in dairy and beef cattle. The initial impression is that growth hormone has value in problem cows such as repeat breeders, cows over 100 days postpartum, and thin cows.

Alison Dixon presented a study conducted at WVa in which the pattern of embryonic and fetal loss in ewes was examined (Item 3F-WVa report). The study was conducted over a two-year period, included the cooperation of nine farms, and entailed the examination of 957 pregnant ewes. The greatest percentage of pregnancy loss in ewes occurred during fetal development (day 45 of gestation to term) rather than late embryonic development (day 25 to day 45 gestation). In addition, the nature of pregnancy losses can be summarized as follows: loss of one or more embryos/fetuses, but not all, of a multiple pregnancy>complete loss of one embryo/fetus in a single pregnancy>complete loss of a multiple pregnancy. These results stand in stark contrast to results from previous studies by Melanie Starbuck in cows, in which the majority of pregnancy loss occurs during the embryonic-fetal transition.

Ron Butler (Cornell) provided updates on projects in progress concerning luteal function (Page 3-Cornell report) and the modification of systemic insulin concentrations with propylene glycol (Page 3-Cornell report). In addition, Ron provided some historical information obtained from Genex, concerning conception rates in cattle from the Northeast over a ten-year period, and encompassing 10,000-15,000 inseminations per month per year (Page 5-Cornell report).

Les Griel (PSU) reported on continuing work at Penn State in which the influence of postpartum uterine bacterial status in dairy cows on ovarian follicular dynamics and fertility has been investigated (Penn State report). Results to date indicate that from Day 15-30 postpartum, most forms of pathogens found within uterine fluid collections consist of A. pyogenes, gram-negative anaerobes, and a small number of other organisms (classified as ‘mixed’). These pathogens, along with coliforms, are associated with a high incidence of services/pregnancy and a high culling percentage. In addition, the Penn State group reports that cows with dominant follicles less than 12 mm in diameter at Day 12 post-insemination have a higher pregnancy rate than cows with follicles grouped into larger diameter categories (Page 2-PSU report).

The group discussed the location and date of next year’s regional meeting. Bob Milvae (UConn) offered to host the meeting for next year. Keith made a motion to accept Bob’s gracious offer. The motion was seconded and voted with unanimous approval.

The next business meeting is scheduled for June 2, 3, and 4th, 2003, at a location to be specified by Bob Milvae at a future date.

Officer elections are as follows:

Dave Townson (UNH) Chair

Keith Inskeep (WVa) Secretary

Bob Milvae (UConn) Director

The meeting adjourned at 4:30 pm.

Respectfully Submitted,

David H. Townson, Secretary