Growth rate and physiology of steers grazing tall fescue inoculated with novel endophytes.

Cattle grazing tall fescue (Festuca arundinacea Schreb.) often develop fescue toxicosis. This condition is thought to be caused by ergot alkaloids produced by the endophyte Neotyphodium coenophialum. Endophytes from wild tall fescue plants, which do not produce ergot alkaloids, were transferred into the endophyte-free tall fescue germplasm, HiMag. The novel associations also lacked the ability to produce ergot alkaloids. Our objective was to determine whether cattle grazing these novel endophyte associations showed signs of fescue toxicosis. At the Fayetteville, Arkansas location, tester steers (n = 72) were assigned to one of four pasture treatments: endophyte-free HiMag tall fescue (HiMag-); 'Kentucky-31' tall fescue infected with its native, toxic endophyte (KY+); and two novel endophyte-infected tall fescue associations, HiMag4 and HiMag9. At the Mount Vernon, Missouri location, steers (n = 54) were used to test three of the four cultivars (HiMag9 was not tested). Ergot alkaloid concentrations in the forage of HiMag4 and HiMag9 were low or undetectable. Respiration rate, rectal temperature, ADG, and hair scores were measured during the grazing period. Blood was collected via jugular venipuncture and used for prolactin, aspartate aminotransferase, alkaline phosphatase (ALP), lactate dehydrogenase (LDH), cholesterol, triglyceride, and creatinine analysis. Weight gains by steers grazing HiMag4 and HiMag9 did not differ from those of steers grazing HiMag-, but were greater than gains (P < 0.05) by steers on the KY+ treatment. Steers grazing KY+ had higher (P < 0.05) respiration rates, rectal temperatures, and hair scores than did steers grazing novel endophyte and HiMag- pastures. Prolactin, ALP, cholesterol, LDH, and triglycerides all were suppressed (P < 0.05) in steers grazing KY+ compared with steers grazing novel endophyte and HiMag- pastures. Steers grazing the novel endophyte tall fescues did not suffer from the decreased weight gains and toxicities associated with fescue toxicosis, resulting in enhanced animal production.

Effects of feeding ergovaline on lamb performance in a heat stress environment.

Lambs exposed to a heat-stressed environment (33 degrees C, 50% relative humidity) were used in three experiments to determine whether ergovaline (EV) is the primary toxin involved in fescue toxicosis. The first study evaluated the effects of feeding diets containing increasing levels of endophyte-infected tall fescue seed (E+) and decreasing levels of endophyte-free tall fescue seed (E-). The second and third study evaluated the response to a diet that contained synthetic EV added to an E- diet and the response to a diet containing endophyte-infected ryegrass seed (R+) with an elevated concentration of EV. In Exp. 1, lambs were fed diets of: 1) 10% E- and 0% E+, 2) 5% E- and 5% E+, or 3) 0% E- and 10% E+. Increasing the percentage of E+ in the diet resulted in a linear decrease (P < 0.01) in feed intake (as-fed basis), skin temperature, thermocirculation index (TCI), and serum prolactin. Body weight gain also decreased (P < 0.06). Respiratory rate and core body temperature were not affected by the 5 or 10% E+ diets. In Exp. 2, lambs were fed diets that contained: 1) 10% E-, 2) 10% E- with synthetic EV added at a level equivalent to the 10% E+ diet, or 3) 10% E+. Feed intake (as-fed basis), body weight gain, and skin temperature did not differ for lambs fed the E- and EV diets. The EV diet elicited a decrease (P < 0.05) in TCI and prolactin compared with the E- diet. The TCI for lambs fed EV did not differ (P > 0.10) from the E+ lambs; however, serum prolactin was lower (P < 0.05) for lambs on the E+ diet than for those fed EV. Core body temperature was not affected (P > 0.10) by feeding EV or E+ fescue seed in Exp. 2. In Exp. 3, lambs were fed diets that contained: 1) 10% E-, 2) 3.24% R+ and 6.76% E-, which added an equivalent amount of EV to E+ diets but reduced concentrations of other ergot alkaloids, or 3) 10% E+. Lambs fed the E+ diet and maintained at 33 degrees C had lowered feed intake (as-fed basis), skin temperature, and TCI compared with lambs fed the E- or R+ diets (P < 0.05). Lambs fed the E+ diet had increased rectal temperatures and lowered serum prolactin compared with lambs on the R+ diet (P < 0.05). Lambs on the R+ diet had a greater rectal temperature and lower serum prolactin than lambs on the E- diet (P < 0.05). These results suggest that EV is a fescue toxin; however, other alkaloids might work synergistically with EV, causing the full expression of fescue toxicosis.

Use of a rat model to evaluate tall fescue seed infected with introduced strains of Neotyphodium coenophialum.

Experimental cultivars of the pasture grass tall fescue are infected with unique strains of the fungal endophyte Neotyphodium coenophialum, which produce low concentrations of ergot alkaloids. A rat model was evaluated as a tool for rapid, initial screening of experimental cultivars considered to be nontoxic. Rats were fed diets that included seed from experimental cultivars of tall fescue with introduced strains of N. coenophialum and a toxic control diet containing seed of the cultivar Kentucky 31 (KY31), with its endemic strain of N. coenophialum. Rats were preconditioned to a nontoxic diet and then fed treatment diets for 13 days with 5 days at thermoneutrality (21 degrees C) followed by 8 days under heat stress (31 degrees C). For most of the 13-day treatment period, rats fed KY31 exhibited depressed daily intake compared to those fed diets of cultivars with introduced endophytes (P < 0.05). In addition, rats fed KY31 exhibited significantly less weight than rats on other diets after heat treatment was imposed. For all initial trials and repeated trials, total intake and total gain calculated at the end of each trial were the most consistent indicators of toxicity.

Reproductive responses to grazing endophyte-infected tall fescue by postpartum beef cows.

The objective was to determine pregnancy rate and stage of embryonic loss in response to grazing endophyte-free (E-; n = 20) or infected (E+; n = 30) tall fescue in postpartum beef cows with calves. Three weeks before estrus synchronization, cow-calf pairs were introduced to pastures (April 1999). Cows were synchronized and bred by AI after detected estrus for a period of 6 d and then by natural service for 62 d. Bulls were rotated weekly to minimize effects of fescue toxicosis on male fertility. Fetal development was monitored weekly between 30 and 60 d of pregnancy and at weaning using transrectal ultrasound. Respiration rate (52.0 +/- 1.4 vs 46.6 breaths/min; P < 0.02) and rectal temperature (39.6 +/- .09 vs 38.8 +/- .12 degrees C; P < 0.001) increased in E+ cows and serum concentrations of prolactin (7.2 vs 57.4 +/- 4.4 ng/mL; P < 0.001), total cholesterol (123.2 vs 149.6 +/- 3.6 mg/dL; P < 0.001), body condition (3.8 vs 5.2 +/- 0.15; P < 0.001; 1 = thin, 9 = fat) and adjusted weaning weight of calves (195.8 vs 210.8 +/- 4.5 kg; P < 0.02) were reduced compared to that of E- cows. Differences were not detected (E- vs E+) for estrus detection rate (84.9 +/- 10.6% vs 80.2 +/- 8.4%), pregnancy rate to synchronized estrus (41.7 +/- 11.8% vs 46.8 +/- 9.5%), overall pregnancy rate 30 d postbreeding (93.8 +/- 6.2% vs 93.5 +/- 5.1%), overall pregnancy rate at 60 d postbreeding (86.7 +/- 10.1% vs 81.2 +/- 8.3%), or serum concentrations of progesterone on day of PGF2alpha treatment (4.5 +/- 0.7 vs 4.5 +/- 0.8 ng/mL). Pregnancy losses that occurred between 30 and 60 d gestation were 6.0 (E-) vs 15.0 (E+) +/- 8.0% (P > 0.10) and occurred after environmental temperatures rose above 37.8 degrees C for three weeks. Total pregnancy losses that occurred by weaning (between 70 and 126 d of gestation) were 5.5 (E-) vs 17.6 (E+) +/- 8.0% (P > 0.10). Pregnancy rate and embryonic losses were not different between cows grazing E- and E+ tall fescue under these management conditions.

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.

Influence of protein supplementation and implant status on alleviating fescue toxicosis.

Heat stress is a major problem in transporting stocker calves with symptoms of fescue toxicosis. Removing calves from tall fescue pastures and offering diets devoid of endophyte-infected tall fescue could reduce the severity of toxicosis and precondition calves for transport to the feedlot. In the present experiment, a pasture phase was used to condition yearling steers to grazing tall fescue and induce symptoms of fescue toxicosis, and a pen phase followed to determine effects of implanting at the start of grazing and protein supplementation (hay only vs hay plus supplement) on short-term changes in rectal temperature and serum prolactin concentration. Neither implant status nor protein supplementation affected (P > 0.10) white blood cell count or rectal temperature. White blood cell counts at the conclusion of the pasture phase averaged 8,778 cells/microL and were within a range indicating no immunological response. Changes in rectal temperature and serum prolactin concentration during the pen phase were not influenced (P > 0.10) by implanting or supplementation. Initial rectal temperatures for the pen phase were high (39.9 degrees C) but declined linearly (P < 0.001) over the first 106 h and were below a normal temperature (39.2 degrees C) by 82 h following removal from tall fescue pastures. Serum prolactin gradually increased (P < 0.001) to a peak by 82 h and stabilized thereafter. Results indicate that neither supplemental protein nor an estrogenic implant influenced recovery indices of fescue toxicosis, whereas removing calves from tall fescue pastures and excluding dietary tall fescue for 3 to 4 d may alleviate symptoms of fescue toxicosis.

Alkaloid binding and activation of D2 dopamine receptors in cell culture.

Ergot and pyrrolizidine alkaloids, either extracted from endophyte-infected tall fescue, synthesized, or purchased commercially, were evaluated in cultured cells to estimate their binding to the D2 dopamine receptor and subsequent effects on cyclic AMP production in GH4ZR7 cells, transfected with a rat D2 dopamine receptor. Ergopeptide alkaloid (alpha-ergocryptine, bromocryptine, ergotamine tartrate, and ergovaline) inhibition of the binding of the D2-specific radioligand, [3H]YM-09151-2, exhibited inhibition constants (K(I)) in the nanomolar range, whereas dopamine was less potent (micromolar). The lysergic acid amides (ergine and ergonovine) were 1/100th as potent as the ergopeptide alkaloids. Ergovaline and ergotamine tartrate were equally effective in inhibiting vasoactive intestinal peptide (VIP)-stimulated cyclic AMP production, with consistent nanomolar effective concentration (EC50) values. The remaining ergopeptide alkaloids (alpha-ergocryptine and bromocryptine), lysergic acid amides (ergonovine and ergine), and dopamine were 1/100th as potent. Two representative pyrrolizidines, N-formylloline and N-acetylloline, exhibited no binding activity at the D2 dopamine receptor or effects on the cyclic AMP system within the concentration ranges of nanomolar to millimolar. Our results indicate that the commercially available ergot alkaloids ergotamine tartrate and ergonovine may be used interchangeably in the D2 dopamine receptor system to simulate the effects of extracted ergovaline and ergine and to examine responses in receptor binding and the inhibition of cyclic AMP.

Ergovaline binding and activation of D2 dopamine receptors in GH4ZR7 cells.

Ergovaline inhibition of radioligand binding to the D2 dopamine receptor and ergot alkaloid inhibition of vasoactive intestinal peptide (VIP)-stimulated cyclic AMP production in GH4ZR7 cells, stably transfected with a rat D2 dopamine receptor, were evaluated. Ergovaline inhibition of the binding of the D2-specific radioligand, [3H]YM-09151-2, exhibited a KI (inhibition constant) of 6.9 +/- 2.6 nM, whereas dopamine was much less potent (370 +/- 160 nM). Ergot alkaloids were also effective in inhibiting VIP-stimulated cyclic AMP production, with EC50 values for ergovaline, ergonovine, alpha-ergocryptine, ergotamine, and dopamine of 8 +/- 2, 47 +/- 2, 28 +/- 2, 2 +/- 1, and 8 +/- 1 nM, respectively. Inhibition of cyclic AMP production by ergovaline was blocked by the dopamine receptor antagonist, (-)-sulpiride (IC50, 300 +/- 150 nM). Our results indicate that ergot compounds, especially ergovaline, bind to D2 dopamine receptors and elicit second messenger responses similar to that of dopamine. These findings suggest that some of the deleterious effects of consumption of endophyte-infected tall fescue, which contains several ergot alkaloids including ergovaline, may be due to D2 dopamine receptor activation.

Changes in pelvic conformation and peripheral estrone concentrations in pre- and post-partum beef cows.

An experiment was conducted to investigate the temporal relationship of peripheral estrone (E1) concentration to changes in the size of the pelvic opening preceding and immediately following parturition. Twenty-six multiparous beef cows were observed from approximately 50 d prepartum to 7 d postpartum. Blood samples were collected at 7 d intervals preceding calving and at 1, 3 and 7 d following for E1 quantitation. Estimates of pelvic opening area were made at the time of blood sampling. Peripheral E1 concentrations were elevated beginning at approximately 25 d prepartum. Dams bearing male fetuses had greater (P less than 0.01) concentrations of E1 than did dams with female fetuses. Calf birth weight was correlated (r = 0.44, P less than 0.01) with E1 levels from 10 d prepartum through parturition. Postpartum pelvic area was greater for cows giving birth to male calves, with no significant differences for calf birth weights by sex. Correlations were observed between E1 concentration, and pelvic area measured from 50 d prepartum to 7 d postpartum (r = 0.26, P less than 0.01), 10 d prepartum to calving (r = 0.42, P less than 0.01), and from calving to 7 d postpartum (r = 0.33, P less than 0.01). Percentage increase in E1 concentration from 50 d prepartum to calving was significantly correlated (r = 0.75, P less than 0.01) to percentage pelvic area increase over the same period. A correlation also exists between maternal E1 concentrations and fetal sex and pelvic area. In summary, the increased estrogen concentrations in cows with male calves may facilitate pelvic spread, resulting in a larger pelvic opening.

Effects of supplemental Bermuda grass hay or corn on intake, digestion and performance of cattle consuming endophyte-infected fescue.

Effects of supplemental Bermuda grass hay (BG) or ground corn on intake, digestion and performance of cattle consuming endophyte-infected fescue (I) were studied. In Exp. 1, a Latin square study, five growing Holstein steers (158.1 kg) consumed I ad libitum and were offered 0, .3, .6, .9 or 1.2% body weight (BW) of BG daily. Total dry matter (DM) intake rose linearly (P less than .05) with increasing BG, although intake was numerically similar with .6, .9 and 1.2% BW of BG. Digestibility was constant with diet (P greater than .10). Six growing Holstein steers used in Exp. 2, a Latin square with a 2 x 3 factorial arrangement of treatments, ingested I or noninfected (NI) fescue hay ad libitum with 0, .5 or 1.0% BW of ground corn. Total DM intake increased linearly as the level of corn rose (P less than .05). Total intake with I increased more with the first than with the second addition of corn, and the opposite occurred with NI (interaction between fescue infection and the quadratic effect of corn level, P less than .10). Organic matter digested (g/d) was greater for NI than for I and rose linearly with increasing corn ingestion (P less than .05). Ninety-six crossbred beef heifers and steers (184.2 kg avg initial live weight) were used in a 77-d fall grazing experiment (Exp. 3) with a 2 x 3 factorial treatment arrangement. Cattle grazed I or NI paddocks and were given no supplement or .34% BW of BG or .65% BW of ground corn on a daily basis (DM).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of frequency of offering and type of supplemental forage on intake and digestion in calves consuming endophyte-infected fescue hay.

Two experiments were conducted to determine the effects of frequency of offering and type of supplemental forage on intake and digestion in calves consuming endophyte-infected fescue hay (I). In Exp. 1, five Holstein steers, averaging 128 kg body weight (BW), were used in a 5 X 5 Latin square experiment. All steers were given free access to I in the afternoon. Morning meals consisted of 1) ad libitum access to I daily (control), 2) .5% BW of Bermuda-grass hay (BG) daily, 3) 1.0% BW of BG every 2nd d, 4) 1.5% BW of BG every 3rd d and 5) 2.0% BW of BG every 4th d. Steers receiving BG consumed less (P less than .01) I and more (P less than .01) total dry matter (DM) than did steers given I alone. Within treatment, I intake was similar (P greater than .10) among days of the feeding cycle without BG. Organic matter (OM) digestion was lower (P less than .05) with than without BG. In Exp. 2, 12 beef calves (Angus and Hereford X Angus; 6 mo of age, 155 kg initial BW) were used in a completely randomized-design experiment. Calves were given ad libitum access to I daily (control) or to BG or wheat hay (WH) on d 1 and I the following 3 d. Hay (I, BG or WH) intake d 1 of the feeding cycle was higher for BG and WH than for the control treatment (16 and 45%, respectively) and higher for WH than BG (25%; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Intake, digestion, passage rate and serum prolactin in growing dairy steers fed endophyte-infected fescue with noninfected fescue, clover or wheat straw.

Growing Holstein steers were used in two Latin-square experiments to determine the effects of supplementation of endophyte-infected fescue hay diets with other forages on intake, digestion, passage rate and serum prolactin concentration. In Exp. 1, five steers (average weight of 186 kg) were fed ad libitum amounts of endophyte-infected and noninfected fescue hays (I and NI, respectively) of similar quality in 0:1, 1:3, 1:1, 3:1 and 1:0 proportions. Total dry matter (DM) intake as a percentage of body weight (BW) linearly decreased .0055% for each 1% increase in dietary I (P less than .05). Dry matter intakes with 100% I and 100% NI diets were 2.13 and 2.72% of BW, respectively. Total tract digestion of neutral detergent fiber (NDF) increased linearly (P less than .05) with increasing I (66.0, 65.9, 66.3, 68.1 and 69.6%). Ruminal passage rate of particulates changed linearly (P less than .05) and quadratically (P less than .10) as I in the diet increased (3.5, 3.4, 2.6, 2.8 and 2.8%/h), while serum prolactin concentration and rectal temperature decreased linearly (P less than .05). In Exp. 2, four steers (average weight of 137 kg) were given ad libitum amounts of wheat straw (WS) or .73% of BW of clover hay (C) at 0800 and free access to either I or NI at 1600. Total intake as a percentage of BW was greatest for C with NI (3.04), intermediate for WS with NI (2.70) and lowest for C with I (2.30) and WS with I (2.23; P less than .05). Fescue intake (percentage of BW) was lowest (P less than .05) for C with I (1.56) and higher (P less than .05) for WS with NI (2.63) than WS with I (2.12); fescue intake for C with NI (2.33) was intermediate (P greater than .10) to WS with NI and WS with I. The results are interpreted to indicate that increasing the dietary level of I depressed intake linearly and markedly. Intake of diets high in I appears to be lower than can be explained only by ruminal-fill factors. When animals that are consuming basal I diets are provided access to nontoxic, high-quality forage, changes in intake may differ from those with basal diets of nontoxic forage.

Effects of offering different amounts and types of supplemental feeds to growing dairy steers fed endophyte-infected fescue hay ad libitum on intake, digestion, passage rate and serum prolactin concentration.

Ten Holstein steers (141 kg) were used in two 5 X 5 Latin-square experiments conducted simultaneously to determine the effects of offering different levels and types of feeds with endophyte-infected fescue given ad libitum. In Exp. 1, steers were given ad libitum access to infected fescue hay in the afternoon; in the morning fescue was given ad libitum (basal) or bermudagrass or clover hays were fed at .5 or 1.0% of body weight (BW). Supplementation did not affect total dry matter intake (P greater than .10), but supplementation at 1.0% of BW yielded total intake greater than supplementation at .5% of BW (P less than .05). Supplementation did not change digestibilities of dry or organic matter (P greater than .10). Particulate passage rate was greater (P less than .10) with supplementation at 1.0 than at .5% of BW, and increasing the level of supplementation from .5 to 1.0% of BW affected fluid passage rate positively with clover but negatively with bermudagrass (interaction, P less than .05). Serum prolactin increased (P less than .05) with all supplementation treatments, although no differences were observed between supplement type-supplementation level combinations (P greater than .10). Ground corn and wheat hay were supplements in Exp. 2. Total intake of dry matter was greater with supplements provided at 1.0 rather than at .5% of BW and for corn rather than wheat hay (P less than .05). Neutral detergent fiber digestion (percent of intake and grams per day) rose when wheat hay was offered at 1.0 vs .5% of BW but declined when the level of supplemental corn increased from .5 to 1.0% of BW (interaction, P less than .05). There were no differences among diets in particulate and fluid passage rates and serum prolactin concentration. Supplementation with nontoxic forage of a basal diet of infected fescue yielded intake substitution when forage was offered at .5% of BW, although incomplete substitution occurred with 1.0% of BW of supplemental forage such that total intake increased as compared to the lower level of supplementation.

Effects of induced fescue toxicosis on plasma and tissue catecholamine concentrations in sheep.

Sixteen crossbred wethers were distributed among four treatments and fed a control ration based on annual rye-orchardgrass (R-O) for 8 days. Indwelling jugular cannulae were installed and experimental regimes begun the following day (experimental day 1). One-half of the wethers were fed a ration based on endophyte-infected Kentucky-31 fescue while the remainder continued to receive the R-O control diet for 10 days. Spiperone, a dopamine antogonist, was administered to one-half of the wethers receiving each ration on days 8 and 9. Plasma prolactin (PRL), dopamine (DA), norepinephrine (NE) and epinephrine (E) were measured in jugular venous blood on days 1, 3, 5 and 7-10 of the trial. On day 10, the animals were decapitated; and DA, NE, E and 3,4-dihydroxyphenylacetic acid (DOPAC) and monoamine oxidase (MAO) were determined in hypothalamic and pituitary tissue. Plasma DA was elevated (P less than .05) following day 8 in wethers fed infected fescue over those fed (R-O), while plasma PRL was reduced (P = .08). Wethers receiving Spiperone had lowered (P less than .05) plasma DA and elevated (P less than .01) plasma PRL. Plasma DA was negatively correlated (P less than .01) with plasma PRL (r = -0.50) following day 8. Plasma NE and E levels and NE, E, DA and DOPAC tissue concentrations were not affected by Spiperone administration or diet. MAO levels in pituitaries were higher (P less than .01) for Spiperone-treated wethers. Wethers receiving the toxic fescue ration exhibited elevated plasma DA concentrations which was associated with depressed prolactin secretion in wethers.(ABSTRACT TRUNCATED AT 250 WORDS)

Fructose utilization for nucleic acid synthesis in the fetal pig.

Eight fetal pigs, in utero, were injected ip with 20 microCi/fetus [U14C]-fructose between d 55 and 65 pregnancy. The isotope was allowed to equilibrate between blood and tissues within injected fetuses for a period of 240 min. Fetal pigs were then sacrificed and nucleic acids were extracted from cold tissue homogenates of skeletal muscle and liver. Nuclide disintegrations per minute recovered in extracted DNA and RNA were used to calculate incorporation of labeled C from fructose. The recovery of labeled C per mmol of nucleic acids from skeletal muscle was greater (P less than .05) than that from liver. Relative incorporation of labeled C into skeletal muscle RNA (395.9 pmol/mmol) was greater (P less than .05) than for DNA (189.5 pmol/mmol). The same trend was observed for liver RNA (78.0 pmol/mmol) and DNA (55.6 pmol/mmol), but differences were nonsignificant. These data suggest that at least part of the high concentration of endogenous fructose measured in fetal pigs in utero is involved in synthesis of nucleic acids, thereby providing substrate for anabolic functions necessary for fetal growth and development.

Inhibition of LH release as influenced by LRF infusion.

Four ovariectomized ewes were infused with LRF (10 mug/hr for 20 hr. Jugular vein blood samples were taken at 30 min intervals. After 16 hr of LRF infusion each animal was given a 1 mg iv injection of LRF. At the end of the 20 hr infusion period, the animals were sacrificed and the pituitary removed. Pituitary and plasma samples were analyzed for LH. Plasma LH concentration reached a peak level of 73 ng/ml, 1.5 hr after starting the LRF infusion. A rapid decline in LH concentration occured for the next 3 hr. The plasma LH concentration declined slowly to preinfusion levels over the next 15.5 hr. A small insigificant rise in plasma LH concentration occured when the animals were given 1 mg of LRF after 16 hr of LRF infusion. At the end of the treatment period pituitary LH concentration varied from 369 to 1360 ng/mg. Two of the four ewes had pituitary LH concentrations similar to untreated ovariectomized ewes. The pituitarys' refractoriness to LRF apparently is not due to depletion of pituitary LH.