Influence of endophyte-infected tall fescue on cyclicity, pregnancy rate and early embryonic loss in the mare

The effects of grazing endophyte-infected tall fescue on luteal function, pregnancy rates, and embryonic loss rates were compared between treated mares (n=18) and untreated controls (endophyte-free, n=12). Mares grazing endophyte-infected fescue demonstrated significantly (P<0.01) prolonged luteal function (22.9 vs 15.8 d) than those grazing endophyte-free fescue. Continuous grazing of endophyte-infected fescue resulted in a decreased (P=0.30) per cycle 14-d viable pregnancy rate (14 31 , 45.2%) compared with that of endophyte-free grazing (12 16 , 75.0%). Early embryonic death rates were higher (P=0.20) in the endophyte-infected group (6 20 , 30.0%) than the endophyte-free group (1 13 , 7.7%). Cumulative pregnancy rates after a 60-d breeding period did not differ between the 2 groups. Embryonic development based on mean vesicle height at 14-d was not significantly different between treatment groups for embryos that maintained viability. Embryos that underwent early embryonic death were smaller (P<0.10) at Day-14 than embryos that maintained viability. Mean plasma progesterone concentrations were significantly (P< 0.01) greater at Day-21 postovulation in endophyte-infected mares in which the embryo remained viable (15.8 ng/ml) than in endophyte-free mares that experienced early embryonic death (9.8 ng/ml) or that demonstrated prolongation of luteal function (11.2 ng/ml). The results of this study suggest that grazing endophyte-infected tall fescue can have a detrimental effect on reproductive efficiency in the mare due to an increase in cycles bred per pregnancy rate, increased early embryonic death rate and prolongation of luteal function.     

Interaction of endophyte-infected fescue and heat stress on ovarian function in the beef heifer.

The objective of the experiment was to examine the interaction of endophyte-infected tall fescue and environmental temperature on follicular and luteal development and function in beef heifers. Heifers were fed endophyte-free or endophyte-infected tall fescue seed at thermoneutral or heat stress temperatures (n = 6/treatment) 4 wk before and 3 wk after synchronized ovulation. All heifers were subjected to thermoneutral conditions (19 degrees C, 50% relative humidity) from Days -7 to -2; temperature increased incrementally from Days -1 to 0 and cycled between 25 degrees C and 31 degrees C between Days 1 and 20 for heat-stressed heifers. Serum was collected and ovaries monitored every other day after induced luteolysis between Days 1 and 23 or until ovulation. Size and location of follicles >4 mm and corpora lutea were recorded. Serum concentrations of prolactin were reduced in heat-stressed heifers fed infected seed and both heat stress and infected seed decreased total cholesterol. Rectal temperature and respiration rate were greatest in heifers fed the infected seed when exposed to maximal temperatures. Heat stress led to reduced diameter of the corpus luteum and serum progesterone compared with thermoneutral conditions. Progesterone was reduced more so in heifers fed infected seed. The combination of infected seed and heat stress was associated with reduced diameter of the preovulatory dominant follicle, and consumption of infected seed led to fewer large follicles during the estrous cycle. Both stressors led to reduced serum estradiol. Impaired follicle function may explain reduced pregnancy rates commonly observed in heifers grazing infected tall fescue pasture.     

Growth and ovarian function of weanling and yearling beef heifers grazing endophyte-infected tall fescue pastures

Growth and ovarian function of crossbred beef heifers grazing low and high endophyte-infected tall fescue pastures were studied for 2 successive years. In April of each year, 20 weanling and 20 yearling heifers were included in the study. All heifers were weighed at 28-d intervals for 112 d. Blood samples were collected from each heifer on Day 0 and +7 of each of five 28-d periods and analyzed for progesterone concentration. Heifers with progesterone concentrations >/= 1.5 ng/ml on either or both Day 0 and +7 were classified as having normal cyclic ovaries. High endophyte-infected fescue pastures adversely altered the ovarian activity (P < 0.05) of weanling heifers in both years. In each trial, average weight gains were lower (P < 0.05) in yearling and weanling heifers grazing the high endophyte-infected pastures than in heifers grazing low endophyte-infected pastures. In 1992, heifers were synchronized with PGF(2alpha) administered on Days 101 and 112. Blood samples were collected on 0, 4, 8 and 12 d after the second PGF(2alpha) injection for progesterone analysis. Heifers grazing high and low endophyte-infected pastures were pastured separately with 4 bulls each and were given heatmount detectors. At 96 h, less estrus activity was observed (P < 0.10; power=0.63) in weanling heifers grazing the high vs. low endophyte pastures although pregnancy rates were similar for all groups. Progesterone concentrations suggested that weanling heifers on the high endophyte pastures had a higher incidence of luteal dysfunction after PGF(2alpha) synchronization. In summary, high endophyte-infected pastures decreased growth in both weanling and yearling heifers, ovarian activity and luteal function were adversely altered in weanling heifers with subsequent decreased estrus response to estrus synchronization.     

The effect of feeding tall fescue seed infected by Acremonium coenophialum on pregnancy and parturition in female rats

1. Female Sprague-Dawley rats (Rattus norvegicus) were randomly assigned to various dietary treatments containing: (1) 100% Purina rodent chow, ad libitum; (2) same as 1, but restricted to daily intake of 7; (3) 50% rodent chow (w/w) and 50% endophyte-free tall fescue (Festuca arundinacea) seed; (4) same as 3, but restricted to intake of 5; (5) 50% rodent chow, 25% endophyte-free tall fescue seed and 25% endophyte-infected (Acremonium coenophialum) tall fescue seed; (6) 50% rodent chow, 12.5% endophyte-free and 37.5% endophyte-infected tall fescue seed; and (7) 50% rodent chow and 50% endophyte infected tall fescue seed. 2. Average daily feed intakes and average daily weight gains decreased with higher levels of endophyte infected seed. 3. Frequency of litter production was affected by all endophyte-infected containing diets. 4. Conception was reduced only in dietary treatment (7). 5. Litter weights, number of pups per litter and weight per pup were proportionally reduced as higher levels of infected seed were incorporated in the ingested diets.     

Short-term mode of secretion of equine chorionic gonadotrop in and the effect of GNRH

Five mature Quarterhorse mares were bled every 30 min for 25 h on day 50 of pregnancy to determine the short-term mode of secretion of equine chorionic gonadotropin (eCG). Three other mares with persistent endometrial cups after abortion were administered gonadotropin releasing hormone (GnRH; 1.0 mug/kg of body weight) and were bled immediately prior to and at 15, 30, 45, 60, 90, 120, 180 and 240 min after GnRH. Concentrations of eCG in plasma of pregnant mares were constant over the 24-h period; the variation of each mare's individual values was no greater (P>.05) than the predicted random variation of the radioimmunoassay. Administration of GnRH had no significant effect on eCG concentrations of mares with persistent endometrial cups over a 4-h period. These data are consistent with a model of eCG secretion in the pregnant mare in which there is little short-term regulation of secretion other than the factors which affect the number of healthy endometrial cup cells within the uterus.     

Endophyte-infected tall fescue diet alters gene expression in heifer luteal tissue as revealed by interspecies microarray analysis

Cattle consuming endophyte-infected tall fescue grass have an associated reduction in circulating progesterone and reduced reproductive rates. In this study, commercially available rat microarrays were used to analyze the gene expression in luteal tissues from heifers fed endophyte-free fescue, endophyte-infected fescue, or endophyte-infected fescue supplemented with the dopamine (DA) antagonist, domperidone. The number of hybridized spots represented approximately 40% of the total 10,000 rat genes/ESTs evaluated. Each luteal sample was analyzed in triplicate, resulting in within treatment correlation coefficients of >/=0.98. Median values of mRNA abundance from luteal tissue taken from the endophyte-infected fed heifers revealed 598 genes and ESTs that were down regulated and 56 genes and ESTs that were upregulated compared with luteal mRNA values from the endophyte-free treatment. There were fewer comparative differences between median values from luteal mRNA from the endophyte-free versus feeding endophyte-infected plus domperidone treated heifers. Only 19 genes and ESTs were upregulated and two were down-regulated.     

Urinary eCG patterns in the mare during pregnancy

Blood and urine samples collected from 12 mares at frequent intervals from 25 to 210 d of pregnancy were analyzed for equine chorionic gonadotropin (eCG). Blood and urine samples were collected daily through two consecutive ovulatory periods from five cyclic mares for comparative purposes. Separate radioimmunoassays (RIA) were developed to detect eCG in the urine and plasma. A simple and quick commercial dipstick enzyme-linked immunospecific assay (ELISA), developed for eCG in the blood, was also utilized in this study to detect eCG in the urine. In the 12 pregnant mares, eCG concentrations in both the plasma and urine as detected by RIA rose significantly on Day 40, peaked by Day 60 and slowly dropped to low levels by Day 200. The dipstick ELISA appeared more reliable for eCG in the plasma than in the urine of the five pregnant mares tested. However, on peak days (50 to 60), both the plasma and urine tested positive in all five mares. Similar eCG profiles were observed when urine samples from seven of the mares were assayed in the dipstick ELISA and RIA. The highest percentage of mares (86%) were positive for eCG by ELISA between Days 65 and 85. The highest concentration of eCG in the urine as detected by RIA was observed between Days 55 and 90. ECG-like immunoactivity was not detected by the ELISA in the urine of cyclic mares, but the RIA showed variable patterns with increases in immunoactivity that could not be correlated with physiological events. In summary, eCG in urine follows a similar profile as the eCG in plasma of mares during their first trimester of pregnancy.     

Connecting plant–microbial interactions above and belowground: a fungal endophyte affects decomposition

Mutualisms can strongly affect the structure of communities, but their influence on ecosystem processes is not well resolved. Here we show that a plant–microbial mutualism affects the rate of leaf litter decomposition using the widespread interaction between tall fescue grass (Lolium arundinaceum) and the fungal endophyte Neotyphodium coenophialum. In grasses, fungal endophytes live symbiotically in the aboveground tissues, where the fungi gain protection and nutrients from their host and often protect host plants from biotic and abiotic stress. In a field experiment, decomposition rate depended on a complex interaction between the litter source (collected from endophyte-infected or endophyte-free plots), the decomposition microenvironment (endophyte-infected or endophyte-free plots), and the presence of mesoinvertebrates (manipulated by the mesh size of litter bags). Over all treatments, decomposition was slower for endophyte-infected fescue litter than for endophyte-free litter. When mesoinvertebrates were excluded using fine mesh and litter was placed in a microenvironment with the endophyte, the difference between endophyte-infected and endophyte-free litter was strongest. In the presence of mesoinvertebrates, endophyte-infected litter decomposed faster in microenvironments with the endophyte than in microenvironments lacking the endophyte, suggesting that plots differ in the detritivore assemblage. Indeed, the presence of the endophyte in plots shifted the composition of Collembola, with more Hypogastruridae in the presence of the endophyte and more Isotomidae in endophyte-free plots. In a separate outdoor pot experiment, we did not find strong effects of the litter source or the soil microbial/microinvertebrate community on decomposition, which may reflect differences between pot and field conditions or other differences in methodology. Our work is among the first to demonstrate an effect of plant–endophyte mutualisms on ecosystem processes under field conditions.     

Systemic relaxin in pregnant pony mares grazed on endophyte-infected fescue: effects of fluphenazine treatment

Tall fescue is one of the most widely grown forage grasses for horses in the United States. However, it is frequently infected with the endophyte Neotyphodium coenophialum which produces ergot alkaloids that cause severe adverse effects in the pregnant mare. The objectives of this study were to determine the effects of fescue toxicosis and fluphenazine on circulating relaxin in pregnant pony mares and evaluate the usefulness of relaxin as a monitor of treatment efficacy. Twelve mares were maintained on endophyte-infected tall fescue pasture. Group TRT (n = 6), received 25 mg of fluphenazine decanoate (i.m.) on Day 320 of gestation while Group UTRT served as untreated controls. Daily blood samples were collected from Day 300 of gestation until Day 3 post partum and analyzed for plasma relaxin concentrations using a homologous equine radioimmunoassay. Mean gestation lengths were 330 +/- 0.7 and 336.5 +/- 3.2 days for TRT and UTRT mares, respectively (P = 0.07). Mean plasma relaxin concentrations in both groups of mares during the week before treatment (Day 313 to 319) were not different (UTRT, 53.4 +/- 11.3 ng/mL; TRT, 61.4 +/- 9.3 ng/mL). In the week after treatment (Day 320 to 326), mean plasma relaxin tended to be higher (P = 0.1) in TRT mares (66.7 +/- 6.2 ng/mL) when compared with UTRT mares (49.6 +/- 6.6 ng/mL), representing a 17.1 ng/mL difference in circulating relaxin between the two groups. Systemic relaxin during the last week before delivery (days relative to parturition) for UTRT and TRT mares was 45.7 +/- 6.7 and 64.7 +/- 6.4 ng/mL (P = 0.06), respectively. At Day -8 and Day -5 relative to parturition, systemic relaxin in TRT mares was significantly higher (P < 0.05) than in UTRT mares. Three of the six UTRT mares and one TRT mare showed clinical symptoms of fescue toxicosis. In the week before delivery, circulating relaxin in mares with problematic pregnancies (39.9 +/- 7.8 ng/mL) was significantly lower than concentrations measured in mares with normal pregnancies (63.4 +/- 5.4 ng/mL; P = 0.03). Clinical observations suggest that a one-time injection with fluphenazine improved pregnancy outcome by reducing the adverse effects of fescue toxicosis concomitant with a stabilization of plasma relaxin concentrations. These data support the hypothesis that systemic relaxin may be a useful biochemical means of monitoring placental function and treatment efficacy in the mare.     

Testosterone secretion during early pregnancy in mares

We have characterized the testosterone secretion pattern during the first 80 d of pregnancy in mares and determined the sources that contribute to circulating testosterone levels during this period. Ten untreated, pregnant mares (Group 1), 10 altrenogest-treated, pregnant mares (Group 2), and 10 altrenogest-treated, pregnant mares in which the CL was eliminated by administration of PGF-2alpha on Day 16 (Group 3) were used in this study. Complete luteolysis occurred following PGF-2alpha administration in all mares in Group 3. Six of the 10 mares in Group 3 did not have an active CL until after Day 60 of pregnancy (Group 3a) and were included in the analysis. The remaining four mares developed a new CL on Days 32, 40, 43 and 49 of pregnancy and were excluded from analysis. Mares without a functional CL (Group 3a) had significantly lower testosterone concentrations than mares with a functional CL (Groups 1 and 2), during the period before equine chorionic gonadotropin (eCG) secretion. At the onset of eCG secretion, testosterone concentrations increased rapidly but the rate of increase decreased with time in mares with a functional CL (Groups 1 and 2). In mares without a functional CL (Group 3a), testosterone concentrations did not increase at the onset of eCG secretion but increased at a gradually increasing rate after Day 50. The lower testosterone concentration in mares without a functional CL before eCG secretion suggests that the CL contributes significantly to the circulating testosterone concentration during the period before eCG secretion. The close time relationship between the onset of eCG secretion and the increase in testosterone secretion in mares with a functional CL and the lack of a testosterone increase in pregnant mares without a functional CL suggest that the increase in testosterone secretion after Day 35 of pregnancy is the result of eCG-stimulated, luteal testosterone synthesis.     

A preliminary study of growth, serum prolactin and reproductive performance of beef heifers grazing Acremonium coenophialum -infected tall fescue

In an experiment with a 2x2 factorial arrangement of treatments, 32 virgin Angus heifers were used to study effects of tall fescue endophyte Acremonium coenophialum and energy supplementation on reproductive performance. Treatments were feeding of endophyte-free (EF) or endophyte-infected (EI) tall fescue without (0%) and with energy supplementation at one percent group mean body weight (1%). Blood samples were collected at 28-day intervals for prolactin (PRL). Starting 21 days before the breeding season, blood was also collected weekly for progesterone (P(4)) and PRL. After 99 days of grazing, heifers received Syncro-Mate-B (SMB) and were then artificially inseminated over a 30-day period. Heifers on EI-0% had a lower (P<0.05) average final body weight. Highest average daily gain was observed in heifers fed EF-1% (0.98 kg/day), whereas, heifers fed EI-0% had lowest average daily gain (0.53 kg/day; P<0.05). Endophyte-infected fescue depressed serum PRL concentrations (171 and 143 ng/ml vs 70 and 85 ng/ml; EF-0% and EF-1% vs EI-0% and EI-1%, respectively; P<0.05). No differences were observed among treatments in luteal activity before or after SMB, in pregnancy rates after a synchronized estrus or in overall pregnancy rates during the 30-day breeding season. Endophyte-infected fescue decreased average daily gain and depressed serum concentrations of prolactin, but had no effect on reproductive performance as measured in this study.     

Prevalence of Escherichia coli O157:H7 and Salmonella in beef steers consuming different forage diets

AIMS: To compare the prevalence of faecal shedding of Escherichia coli O157:H7 and Salmonella in growing beef cattle consuming various forages. METHODS AND RESULTS: In Experiment I, faecal samples were collected from steers grazing either endophyte-infected (E+) tall fescue or common bermudagrass (CB). Steers grazing E+ tall fescue were confined to a dry-lot pen and fed CB hay ad libitum for 10 days. In Exp. II, faecal samples were collected from steers grazing either E+ or novel endophyte-infected (NE) tall fescue and treated with one of two anthelmintics: ivermectin (I) or fenbendazole (F). In Exp. I, prevalence of E. coli O157:H7 was less in E+ tall fescue steers fed CB hay than steers grazing CB. More I-treated steers shed Salmonella than F-treated steers at 42-day postanthelmintic treatment but shedding of Salmonella was similar between anthelmintics at day 63 in Exp. II. CONCLUSIONS: Faecal shedding of pathogenic bacteria was not affected by grazing E+ tall fescue. Alterations of forage diets may influence the prevalence of E. coli O157:H7, and anthelmintic treatment could affect faecal shedding of Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of factors that influence shedding of pathogenic bacteria in cattle is necessary to develop on-farm intervention strategies aimed at reducing pathogen shedding.     

The Effects of Endophytes on Seed Production and Seed Predation of Tall Fescue and Meadow Fescue

Fungal endophytes of grasses are often included in agricultural management and in ecological studies of natural grass populations. In European agriculture and ecological studies, however, grass endophytes are largely ignored. In this study, we determined endophyte infection frequencies of 13 European cultivars and 49 wild tall fescue (Schedonorus phoenix) populations in Northern Europe. We then examined seed production and seed predation of endophyte-infected (E+) and endophyte-free (E?) tall fescue (in wild grass populations and in a field experiment) and meadow fescue (Schedonorus pratensis; in a field experiment only). Endophytes were detected in only one of the 13 cultivars. In contrast, >90% of wild tall fescue plants harbored endophytes in 45 wild populations but were absent in three inland populations in Estonia. In three wild tall fescue study sites, 17%, 22%, and 56% of the seeds were preyed upon by the cocksfoot moth. Endophyte infection did not affect seed mass of tall fescue in the field experiment. However, seed predation was lower in E+ than E? grasses in the two tall fescue populations with higher predation rates. For meadow fescue, the mean number of seeds from E+ plants was higher than E? plants, but E? and E+ seeds had equal rates of predation by the moth. Our results suggest that the effects of grass endophytes on seed production and cocksfoot moth seed predation vary considerably among grass species, and the effects may depend on herbivore pressure and other environmental conditions.     

Endophytic fungus improves growth and metal uptake of Lolium arundinaceum Darbyshire ex. Schreb

The effect of endophyte infection on plant growth, cadmium (Cd) uptake, and Cd translocation was investigated using tall fescue (Lolium arundinaceum) grown in greenhouses in contaminated solution. Endophyte infection significantly increased tiller number and biomass of the host grass under both control and Cd-stress conditions. Endophyte infection not only enhanced Cd accumulation in tall fescue, but also improved Cd transport from the root to the shoot. Under 20 mg L(-1) Cd stress, the phytoextraction efficiency of endophyte-infected (EI) tall fescue was 2.41-fold higher than endophyte-free plants. Although the total Cd accumulation in EI tall fescue was insufficient for practical phytoextraction applications, the observed high biomass production and tolerance of stress from abiotic factors including heavy metals, gives endophyte/plant associations the potential to be a model for endophyte-assisted phytoremediation of metal-polluted soils.     

Assessing short-term responses of prokaryotic communities in bulk and rhizosphere soils to tall fescue endophyte infection

In contrast to endophyte-free (E−) tall fescue, endophyte-infected (E+) tall fescue pastures appear to enhance soil carbon sequestration. A hypothetical mechanism that may account for the enhanced carbon sequestration is that the E+ tall fescue affects the soil microbial community or components of it that are involved in organic carbon turnover. A 60-week mesocosm study with a factorial arrangement of soil type, loamy sand (LS) and clay loam (CL), and E+ and E− tall fescue was conducted to determine if the soil microbial communities were affected by the presence of the endophyte. Bulk and rhizosphere soil samples were fixed in paraformaldehyde, and prepared for total direct microbial counts, and with a combination of one of a domain or subdivision fluorescent oligonucleotide probe for enumerating metabolically active Eubacteria, bacterial subdivisions, and Archaea. E+ tall fescue suppressed the archaeal and high G+C gram-positive bacterial communities of the bulk CL, the delta-proteobacterial community in the rhizosphere CL, and the Planctomycetes community of the rhizosphere LS. In the long-term, suppression of these microbial communities may be a factor in enhanced soil carbon sequestration associated with E+ tall fescue.     

Antagonistic effects of simultaneous exposure of ergot alkaloids on kidney adenosine triphosphatase system

Much of the research on fescue toxicosis has concentrated on evaluating animal response to grazing endophyte-infected (E+) versus endophyte-free tall fescue or the effects of single toxins such as ergonovine (EN), ergovaline (EV), or ergotamine (ET) on animal performance. Such approaches have eliminated the opportunity to test the possible additive, synergistic, or antagonistic interactions of one or more ergot alkaloids with the other ergot alkaloids found in E+ tall fescue. This study was conducted to determine the effects of simultaneous exposure of pairs of EN, EV, and ET on the kidney adenosine triphosphatase (ATPase) system in vitro. Tests were performed using three separate rat kidney homogenates and were repeated four times at concentrations of 0, 75, and 200 microM. Individually, EN, EV, and ET induced dose-dependent inhibitions of kidney Na(+)/K(+) ATPase, with EN being most potent, followed by purified EV, and then by ET. The ergot alkaloids inhibited Mg(2+) ATPase to a lesser degree than Na(+)/K(+) ATPase, with EN again being the most potent toxin. Simultaneous exposure to any combination of the ergot alkaloid pairs tested (EV + ET, EV + EN, and ET + EN) resulted in significant interactions (P < 0.05), indicating antagonistic effects on the inhibition of Na(+)/K(+) ATPase and Mg(2+) ATPase for most concentration combinations. These interactions suggest that in studies of the effects of any ergot alkaloid on animal performance, effects of other ergot alkaloids may also be present. Effects may not be additive, as was the case in this study, and the presence of one toxin may enhance or hinder the effectiveness of others.     

Influence of endophyte infection, plant age and harvest interval on Rhopalosiphum padi survival and its relation to quantity of N-formyl and N-acetyl loline in tall fescue

Rhopalosiphum padi L. (Homoptera: Aphididae) is sensitive to loline alkaloids present in tall fescue, Festuca arundinacea Shreb., infected with the endophytic fungus, Acremonium coenophialum Morgan-Jones & Gams. Aphid survival was higher on endophyte-free plants regardless of plant age after germination or age of regrowth tissue after clipping. Survival of aphids on endophyte-infected grass was lower on young tissue but increased as plants aged, although it never reached the same level on endophyte-free plants. Both N-formyl and N-acetyl loline increased as uncut or regrowth tissue aged; however, this was influenced by the age of the plant at the initial cut and the clipping frequency. Although even small amounts of loline cause high aphid mortality, the aphids are able to survive on endophyte-infected plants if the tillers have senescing leaves which contain lower amounts of loline. Preference for senescing leaves may help R. padi avoid plant parts containing high amounts of toxic allelochemicals, thus contributing to higher numbers of aphids on older, endophyte-infected plants.     

Changes in vascular perfusion of the endometrium in association with changes in location of the embryonic vesicle in mares

The equine embryonic vesicle is mobile on Days 12-14 (Day 0 = ovulation), when it is approximately 9-15 mm in diameter. Movement from one uterine horn to another occurs, on average, approximately 0.5 times per hour. Mobility ceases (fixation) on Days 15-17. Transrectal color Doppler ultrasonography was used to study the relationship of embryo mobility (experiment 1) and fixation (experiment 2) to endometrial vascular perfusion. In experiment 1, mares were bred and examined daily from Day 1 to Day 16 and were assigned, retrospectively, to a group in which an embryo was detected (pregnant mares; n = 16) or not detected (n = 8) by Day 12. Endometrial vascularity (scored 1-4, for none to maximal, respectively) did not differ on Days 1-8 between groups or between the sides with and without the corpus luteum. Endometrial vascularity scores were higher (P < 0.05) on Days 12-16 in both horns of pregnant mares compared to mares with no embryo. In pregnant mares, the scores increased (P < 0.05) between Day 10 and Day 12 in the horn with the embryo and were higher (P < 0.05) than scores in the opposite horn on Days 12-15. In experiment 2, 14 pregnant mares were examined from Day 13 to 6 days after fixation. Endometrial vascularity scores and number of colored pixels per cross-section of endometrium were greater (P < 0.05) in the endometrium surrounding the fixed vesicle than in the middle portion of the horn of fixation. Results supported the hypothesis that transient changes in endometrial vascular perfusion accompany the embryonic vesicle as the vesicle changes location during embryo mobility.     

Luteogenic and luteotropic effects of eCG during pregnancy in the mare

The role of eCG during pregnancy was evaluated through the study of the temporal relationships between changes in eCG and progesterone concentrations and the formation of supplementary corpora lutea (SCL) in mares impregnated with donkey semen (mule pregnancies) or with horse semen (equine pregnancies). Concentrations of eCG were higher (p<0.01) in equine than in mule pregnancies between weeks 6.5 and 13. Progesterone concentrations were higher in equine than in mule pregnancies between weeks 9 and 17. All animals developed at least one SCL, but more SCL accumulated during equine pregnancies than during mule pregnancies (1.9 ± 0.2 vs 1.2 ± 0.1; p<0.01). In equine pregnancies, the mares that formed a second SCL had higher eCG concentrations (p<0.05) during the two weeks preceding its formation than those mares remaining with only one SCL. Mares that formed a third SCL had higher (p<0.5) eCG levels than those remaining with one or two SCL. Mares with equine pregnancies that formed three SCL had higher progesterone concentrations (p<0.05) than those that formed only one or two SCL. No differences were found in progesterone or eCG concentrations between mares with mule pregnancies that accumulated different numbers of SCL during pregnancy (p>0.05). It is concluded that eCG stimulates both the development of new SCL and the function of existing CL. While these effects are clearly expressed in mares impregnated by horses, the low eCG concentrations during mule pregnancies reduce the impact of this hormone on CL formation and function.     

Importance of host plant species, Neotyphodium endophyte isolate, and alkaloids on feeding by Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae

Three grass host species--tall fescue, Festuca arundinacea Schreber; meadow fescue, Festuca pratensis Hudson; and perennial ryegrass, Lolium perenne L.--each infected with a number of different Neotyphodium endophyte isolates, were investigated for their effects on fall armyworm, Spodoptera frugiperda (J.E. Smith). Alkaloid profiles varied among associations. Choice and no-choice tests comparing feeding and early development of S. frugiperda larvae on endophyte-infected and endophyte-free leaf blade material were performed. Endophyte-mediated resistance to S. frugiperda was greatest in meadow fescue and weakest in tall fescue. Some endophyte isolates, particularly in perennial ryegrass and meadow fescue, had a major effect on feeding and development of S. frugiperda, whereas others had no effect or were only weakly efficacious. In tall fescue, some associations deterred S. frugiperda from feeding in choice tests but had no effect on development, whereas larvae reared on other associations weighed significantly more than control larvae fed endophyte-free grass. It was concluded that the deleterious consequences of endophyte infection were easily masked by other factors in tall fescue. Relative leaf age had no effect on feeding preferences in the three host species. Chemical analysis of herbage from the plants used, and results from a no-choice study using spiked artificial diets, failed to individually implicate any of the major known alkaloids (peramine, lolitrem B, ergovaline, and lolines) in the observed effects on S. frugiperda. Hypotheses explaining these observations, and their impact on creating desirable grass-endophyte associations for use in pastures, are discussed.