The effect of the endophyte (Acremonium coenophialum) and associated toxin(s) of tall fescue on serum titer response to immunization and spleen cell flow cytometry analysis and response to mitogens

Experiments were conducted with rats and mice to evaluate the effect of the consumption of endophyte (Acremonium coenophialum) and associated toxin(s) infected tall fescue on humoral and cellular aspects of immune function. Treatment diets were: (1) rodent chow (RC) or (2) rodent chow mixed 1:1 (w/w) with endophyte infected (E+) or (3) non-infected (E-) tall fescue seed. Rats fed the E+ diet in experiment 1 (43 days) exhibited a lower (P less than 0.05) serum titer to sheep red blood cell (SRBC) immunization than those fed the E- diet (38.4 vs 131.3). The E+ rats also had lower (P less than 0.01) white cell counts than either RC or E- groups (5225 vs 8959 and 7491/mm3). Spleen cells from mice fed the E+ diet for 37 days exhibited a reduced (P less than 0.05) response to the mitogens Concanavalin A and lipopolysaccharide. Flow cytometric analysis revealed a significant (P less than 0.01) 42% increase in T suppressor cell numbers in spleens of mice fed the E+ vs RC diets.     

Effects of feeding Kentucky 31 tall fescue seed infected with acremonium coenophialum to laboratory rats

An endophytic fungus found in Kentucky 31 (KY-31) tall fescue has been associated with the "summer syndrome" (fescue toxicity) in grazing cattle. Four experiments were conducted to investigate the use of weanling rats as an animal model to study the fescue toxicity syndrome. When graded levels of fungus-infected (INF) or fungus-free (FF) KY-31 seed were fed, feed intake and growth rates were inversely related to the amount of INF seed in the diet. When diets containing 50% seed were fed, rats eating INF seed had reduced (P less than .05) daily water consumption (9 vs 20 ml) and lowered (P less than .05) body temperatures (36.8 vs 37.7 C) when compared with the FF treatments. Autoclaving the seed increased feed intake and growth rate of rats receiving both INF and FF seed diets, but daily gains continued to be less (P less than .05) in the rats fed autoclaved INF diets when compared with rats fed autoclaved FF diets (2.3 vs 6.8 g). Rats fed diets containing either 40% shredded wheat on which the fescue endophyte had been grown or 40% shredded wheat that was not innoculated did not differ in performance. Crude fiber, nitrogen-free extract, gross energy and dry matter digestibilities were enhanced (P less than .05) by the presence of INF seed in the diets when compared with the FF seed treatments. Crude protein and ether extract digestibilities and apparent biological value of fescue protein were not affected by presence of the fungus. The effect of INF seed on organ weights was inconsistent.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of endophyte-infected tall fescue on serum prolactin and progesterone in gravid mares.

Thirty mares in late gestation were used in a 3-yr study to assess effects of the tall fescue endophyte Acremonium coenophialum on serum prolactin (PRL) and progesterone. Two paddocks of each treatment, 0 or 100% infected 'Kentucky 31' tall fescue, were grazed by the mares for 21 d. Blood was collected three times per week until parturition. At 7-d intervals, mares were challenged with thyrotropin-releasing hormone (TRH) while grazing and blood was collected postinjection. Mares grazing 100% infected tall fescue (E+) had decreased serum PRL compared with mares grazing the 0% infected tall fescue (E-) in 2 of 3 yr. Within 8 d postgrazing, serum PRL for E+ mares equaled or surpassed values of the E- mares. Serum PRL was not different during the 3rd yr. In response to TRH, serum PRL rate of increase was similar between treatments but remained elevated (P less than .01) in the E+ mares at the 180-, 240-, and 300-min sample times. Serum progesterone was lowered (P less than .05) by E+ but increased to control values within 10 d postgrazing. It is concluded that serum PRL and progesterone in the gravid mare were decreased by the presence of A. coenophialum in 'Kentucky 31' tall fescue grass but normal levels were reestablished within 2 to 3 wk.     

Actions of isoflurane and halothane in pregnant mares.

Eighteen healthy, pregnant mares scheduled for laparotomy and uterine manipulation were randomly allotted to 2 equal groups. After IV administration of xylazine hydrochloride and thiamylal sodium, general anesthesia was maintained with halothane (HALO) or isoflurane (ISO) in oxygen. Results of cardiovascular measurements were similar with both inhalant anesthetics; mean arterial blood pressure was 79 and 82 mm of Hg with HALO and ISO, respectively. Respiratory rate decreased most with ISO (mean frequency was 4 and 9 breaths/min with ISO and HALO, respectively). Partial pressure of arterial CO2 was increased similarly with HALO and ISO. Partial pressure of arterial O2 varied greatly among mares and decreased with duration of use of both anesthetics. Recovery time from anesthesia was significantly (P < 0.05) shorter after use of ISO vs HALO. Minor superficial injuries were associated with recovery from both anesthetics (in 5 mares with ISO and in 1 mare with HALO). Physical signs of postanesthetic myopathy or vital-organ dysfunction were not associated with either agent.     

Effects of ammoniation of endophyte fungus-infested fescue hay on serum prolactin concentration and rectal temperature in beef cattle

A group of 48 crossbred steers (approx 250 kg each) were used to determine the effects of various diets and treatments on serum prolactin concentrations and rectal temperatures. The steers were randomly assigned to groups fed the following: (1) endophyte fungus-infested fescue hay; (2) ammoniated endophyte fungus-infested fescue hay; (3) endophyte fungus-infested fescue hay plus 1 g of thiabendazole (TBZ)/9.1 kg of body weight at 7-day intervals; (4) ammoniated endophyte fungus-infested fescue hay plus 1 g of TBZ/9.1 kg at 7-day intervals; (5) ground Bermuda grass hay; and (6) endophyte-fungus-free fescue hay. Blood samples for prolactin determinations and rectal temperatures were obtained biweekly beginning on week 3 and continuing through week 9. A significant correlation (P less than 0.05) between low prolactin concentrations and high rectal temperatures in cattle eating endophyte-infested fescue hay was determined; however, as the prolactin concentrations approached normal (control animal) concentrations, this relationship between serum prolactin and rectal temperatures was not observed. Two antifungal agents used in this experiment (thiabendazole and ammonia) appeared to have different effects on the variables measured. Thiabendazole had no significant effect on prolactin concentrations or rectal temperatures of cattle ingesting endophyte-infested fescue hay, whereas ammoniation of this hay induced significantly higher (P less than 0.05) prolactin concentrations and lower rectal temperatures than in steers receiving only endophyte-infested fescue hay. Therefore, ammonia may be valuable not only as an aid in determining the cause of the fescue problem but also as a practical solution to some of the fescue-related economic problems in the cattle industry. Also, prolactin concentrations may be valuable in diagnosing fescue-related problems.     

Performance, forage utilization, and ergovaline consumption by beef cows grazing endophyte fungus-infected tall fescue, endophyte fungus-free tall fescue, or orchardgrass pastures.

Two 120-d trials (May to September, 1988 and 1989) determined the effects of grazing tall fescue (two varieties) or orchardgrass on forage intake and performance by beef cows. Each summer, 48 cow-calf pairs grazed endophyte-infected Kentucky-31 tall fescue (KY-31), endophyte-free Mozark tall fescue (MOZARK), or Hallmark orchardgrass (OG) pastures (16 pairs/treatment). Forage OM intakes and digestibilities were determined during June and August each year. Cow and calf BW and milk production were determined every 28 d. During June of both years, OM intakes did not differ (P greater than .10) among treatments. During August of 1988, intakes were 18% lower (P less than .05) by KY-31 cows (1.6% of BW) than by MOZARK or OG cows (average 1.95% of BW); however, no differences (P greater than .10) were measured in August of 1989. Estimates of ergovaline consumption during June from KY-31 were between 4.2 (1988) and 6.0 mg/d (1989), whereas August estimates were between 1.1 (1988) and 2.8 mg/d (1989). Ergovaline in MOZARK estrusa was below detection limits, except in August of 1989. Cows that grazed KY-31 lost three times (P less than .01) more BW than cows that grazed MOZARK or OG (42 vs 9 and 13 kg, respectively). Milk production by KY-31 cows was 25% lower (P less than .01) than that by cows that grazed MOZARK or OG (6.0 vs average of 8.0 kg/d). Similarly, slower (P less than .01) calf gains were noted for KY-31 than for MOZARK or OG (.72 vs .89 and .88 kg/d, respectively). Cows grazing KY-31 experienced accelerated BW loss and reduced milk production and weaned lighter calves than did cows grazing MOZARK or OG. Decreased performance was not explained by consistently reduced forage intakes; hence, altered nutrient utilization was suspected.     

内生真菌对苇状羊茅和多年生黑麦草影响的研究进展

综述了内生真菌的分类及其在宿主苇状羊茅Festuca arundinacea和多年生黑麦草Lolium perenne中的分布与传播方式,阐述了内生真菌对苇状羊茅和多年生黑麦草生长、抗生物(病虫害)和非生物胁迫(P胁迫和干旱胁迫等)、以及对其他草类的竞争他感作用的影响,并提出今后的研究方向.     

Reproductive traits, lactation and foal growth in mares fed altrenogest

Lactating mares were assigned as controls or fed altrenogest (.044 mg.kg body wt-1.d-1) for 15 d after foaling. Mares (n = 6) fed altrenogest were inseminated during the first estrus after treatment and mares (n = 6) in the control group were inseminated during the second postpartum estrus. Ovulation during the estrus in which mares were inseminated occurred 26 +/- 1 d postpartum for treated mares and 36 +/- 1 d postpartum for control mares. The percentage of mares conceiving was not different for control (67%) and alternogest-treated (100%) mares. No differences were observed in tone and size of the uterus or size of the ovulatory follicle between treated and control groups. Uterine cultures and biopsies collected on d 7 and 15 postpartum were similar between treatment and control groups in bacterial populations or endometrial epithelial cell height. Blood was collected on d 7, 11, 15, 19 and 23 postpartum, and concentrations of estradiol-17 beta in serum were determined by radioimmunoassay. Mean concentrations of estradiol-17 beta across days were 10 +/- .8 and 12 +/- .6 pg/ml for control and treated mares, respectively. Concentrations of serum estradiol-17 beta were higher (P less than .05) in treated mares on d 23 postpartum. Daily milk yields, determined by the weigh-suckle-weigh method, and milk composition were similar between treatment groups on each collection day. Altrenogest can be used to predictably delay estrus in the postpartum mare without altering fertility, yield and composition of milk, or foal growth.     

Influence of dietary endophyte (Neotyphodium coenophialum)-infected tall fescue (Festuca arundinacea) seed on fecal shedding of antibiotic resistance-selected Escherichia coli O157:H7 in ewes

The objectives were to determine the effects of short-term feeding of a toxic endophyte (Neotyphodium coenophialum)-infected tall fescue seed (Festuca arundinacea, cultivar 'Kentucky 31') on fecal shedding and intestinal concentrations of Escherichia coli O157:H7 and the concentrations of prolactin, cortisol, and NEFA in experimentally inoculated ewes. Twelve ewes (mean BW = 46 +/- 2 kg) were fed a diet containing either high endophyte-infected (HI-E) or low endophyte-infected (LO-E) tall fescue seed for 7 d. Each diet consisted of 50% (as-fed basis) tall fescue seed. Ewes were experimentally inoculated with antibiotic resistance-selected E. coli O157:H7 on d 1 of the feeding treatment, and fecal shedding of inoculated pathogens was monitored daily on d 2 to 6. On d 7, ewes were weighed and euthanized, and tissues and contents were sampled from the ileum, cecum, and rectum for quantitative enumeration of E. coli O157:H7. Urine was collected at euthanization to determine total ergot alkaloid concentrations. Ewes fed HI-E had lower (P < 0.001) DMI than did ewes fed LO-E (0.8 and 1.6 +/- 0.1 kg/d of DMI for HI-E and LO-E ewes, respectively); consequently, there was a tendency (P = 0.06) for HI-E ewes to lose 0.3 +/- 0.4 kg of BW/d and LO-E ewes to gain 0.2 +/- 0.4 kg of BW/d during the 7 d. Urinary ergot alkaloids were increased (P < 0.001) in ewes fed HI-E (47.8 +/- 9.4 ng/mg of creatinine) compared with those fed LO-E (6.2 +/- 9.4 ng/mg of creatinine). Prolactin tended (P = 0.06) to be decreased in ewes fed HI-E (7.2 +/- 7.0 ng/mL) compared with those fed LO-E (27.7 +/- 7.0 ng/mL). Fecal shedding of E. coli O157:H7 tended (P = 0.06) to be increased in HI-E ewes [5.4 cfu (log10)/g of feces] compared with LO-E ewes [4.5 cfu (log10)/g of feces]. The population of E. coli O157:H7 in luminal contents from the ileum, cecum, and rectum did not differ (P > 0.36) between treatments. Treatment did not influence (P = 0.30) the occurrence of E. coli O157:H7 in cecal or rectal tissues; however, ileal tissues from HI-E ewes tended (P = 0.12) to have an increased incidence of E. coli O157:H7. Concentrations of NEFA tended (P = 0.12) to be greater in HI-E ewes than in LO-E ewes, whereas cortisol was similar (P = 0.49) for HI-E and LO-E ewes. We conclude that short-term feeding of HI-E tall fescue seed may alter the concentrations of prolactin and NEFA, and may increase fecal shedding of E. coli O157:H7 in experimentally inoculated ewes.     

Herniation of the abdominal wall in pregnant mares

Abdominal wall hernia was detected in 4 pregnant mares. Antemortem diagnosis of the specific abdominal wall lesion was difficult. Ventral deviation of the abdomen, associated abdominal wall edema, and pain indicated rupture of the prepubic tendon. Three mares examined at necropsy did not have a rupture of the prepubic tendon but did have herniation of the abdominal wall. Abdominal pain was severe and was compounded by incarceration or entrapment of viscus.     

The effect of excess dietary iodine on pregnant mares and foals.

On a thoroughbred stud four foals were born with greatly enlarged thyroids and leg weakness. Two foals died within 18 hours of birth, the others subsequently recovered. An enlarged thyroid was also evident in one of the resident mares. The thyroids from the dead foals were hyperplastic. Feed analyses showed that the mares had an iodine intake of about 83 mg daily, 8-8 ppm of the dietary dry matter, due almost entirely to the high iodine content of a proprietary compound horse nut which had been fed at the daily rate of 12 lb per head. It was concluded from the histology of the thyroids, the high intake of iodine, the lack of response to treatment with potassium iodide and the elevated levels of serum protein bound iodine that the condition of the foals on the stud was caused by an excess of iodine fed to the mares during pregnancy.     

Growing steers grazing high versus low endophyte (Neotyphodium coenophialum)-infected tall fescue have reduced serum enzymes, increased hepatic glucogenic enzymes, and reduced liver and carcass mass

It is well established that grazing Neotyphodium coenophialum-infected forages results in reduced BW gain and serum prolactin concentrations of cattle. The objective of this study was to determine the potential effects of toxic endophyte-infected tall fescue consumption on blood metabolites, carcass characteristics, and content of proteins critical for AA metabolism in the liver, kidney, and LM tissue of growing steers. Steers grazed a low toxic endophyte (LE; 0.023 microg/g ergot alkaloids) tall fescue-mixed grass pasture (n = 9; BW = 266 +/- 10.9 kg; 5.7 ha) or a high toxic endophyte (HE; 0.746 microg/g of ergot alkaloids) tall fescue pasture (n = 10; BW = 267 +/- 14.5 kg; 5.7 ha) from June 14 through at least September 11 (> or =89 d). No difference was observed for BW (P < 0.10) for the overall 85-d growth period. Also, no differences were observed for ribeye area/100 kg of HCW (P > 0.91), backfat (P > 0.95), or backfat/100 kg of HCW (P > 0.67). However, ADG (P < 0.01), final BW (P < 0.05), HCW (P < 0.01), dressing percentage (P < 0.01), ribeye area (P < 0.01), whole liver wet weight (P < 0.01), and whole liver wet weight/100 kg of end BW (P < 0.01) were greater for LE steers than HE steers. After 85 d of grazing, serum concentrations of alkaline phosphatase (P < 0.05), alanine aminotransferase (P < 0.01), aspartate aminotransferase (P < 0.03), cholesterol (P < 0.01), lactate dehydrogenase (P < 0.01), and prolactin (P < 0.01) were less for HE than LE steers. At slaughter, hepatic content of cytosolic phosphoenolpyruvate carboxykinase (P < 0.01) was greater in HE steers than LE steers. Hepatic content of aspartate aminotransferase (P < 0.01) also was greater, whereas renal and LM content were not (P > or = 0.42). No differences (P > or = 0.15) were observed for hepatic, renal, and LM content of alanine aminotransferase, glutamate dehydrogenase, glutamine synthetase, and 3 glutamate transport proteins. These data indicate that the HE steers displayed classic endophyte toxicity symptoms for growth and blood variables, classic symptoms that were concomitant with novelly identified altered glucogenic capacity of the liver and decreases in carcass characteristics.     

禾草内生真菌检测方法研究进展

从最初发现黑麦草(Lolium perenne)和高羊茅(Festuca arundinacea)引起家畜中毒到后来其携带的内生真菌被发现,以及进一步的研究证实,内生真菌的存在不但导致家畜中毒而且能显著提高宿主在群落中的竞争力,也因禾草内生真菌的这种重要生理和生态作用,使其逐步成为国内外研究的热点之一,这也给内生真菌检测技术的发展提供了契机。初期主要是借鉴病原真菌等微生物检测的一些较成熟的方法,对禾草内生真菌进行染色镜检或分离检测,但容易受宿主种类、物候期、检测部位等因素的影响,造成检测结果的不准确。随着分子生物学、遗传学等学科的快速发展,酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)、聚合酶链式反应法(polymerase chain reaction,PCR)、实时荧光PCR法(realtime PCR)等现代分子技术在禾草内生真菌方面的应用和改进,使得内生真菌的检测方法不断推陈出新,这些方法能在一定程度上对传统检测方法给予有效的补充。快速有效的确定禾草内生真菌的存在、分布规律、分类地位等,需要准确合理地选择特异性较强的检测方法,例如定性或定量检测,并结合经典的染色镜检和分离法对禾草内生真菌进行确定。本研究对以上内容进行了综述,以期更深层次的借鉴和发展关于其它微生物的经典检测方法,同时开发具有禾草内生真菌特异性的检测技术,这些技术不仅能够鉴定内生真菌存在与否,而且能对内生真菌进行定量和活性的检测,是发展的重要方向和热点。对内生真菌定性、定量和活性的检测是未来值得研究的重点。     

Clinical biochemistry of pregnant and nursing mares

BACKGROUND: Pregnancy and lactation result in increased metabolic demands. Although homeostatic mechanisms function to keep substances in blood at relatively constant levels, some changes in the concentrations of routine clinical chemistry analytes are likely to occur. OBJECTIVES: The purpose of this study was to determine what physiological changes occur in serum clinical biochemistry analytes in pregnant and nursing mares, and to determine whether the changes were substantial enough to warrant separate reference intervals for pregnant or lactating horses. METHODS: Forty-two Quarter Horse, Thoroughbred, Saddlebred, Standardbred and Morgan mares were entered into the study while pregnant. They were bled once each month through birth. Studies were continued on 20 mares until their foals were weaned. Test results were tabulated by time before or after birth. Serum biochemistry values were determined by standard methods using automated analyzers. Test results were analyzed using the Kruskal-Wallis 1-way ANOVA on ranks. If a significant difference was found (P<.05), Dunn's multiple comparison procedure was performed on all pairs. Results from pregnant and nursing mares also were compared with a reference group of 19 open, nonlactating mares. RESULTS: Serum triglyceride, potassium, creatinine, and total bilirubin concentrations were lower during lactation than during pregnancy. Serum calcium concentration also was slightly decreased at 2 time points during lactation. Triglyceride concentration was highest during midgestation, while bilirubin and creatinine values increased, and potassium and calcium remained constant during pregnancy. Serum urea concentration also remained constant during pregnancy but increased during lactation. Serum phosphate concentration was lowest during midgestation and highest during lactation. Total CO2 values were highest, and anion gaps were lowest, during midgestation. No significant differences were found in serum albumin, globulin, albumin:globulin ratio, total protein, or glucose values. When compared with the reference group of open mares, serum triglyceride, potassium, bilirubin, and total CO2 concentrations were lower, and anion gap was higher in horses that were nursing. CONCLUSIONS: Although most biochemical values remained relatively constant, significant differences were observed during pregnancy and lactation. Changes in the concentrations of triglycerides, potassium, bilirubin, total CO2, and anion gap during lactation were substantial enough to warrant separate reference intervals for lactating horses.     

Effects of fescue toxicosis on reproduction in livestock.

Fescue toxicosis in livestock is due to ingestion of endophyte (Acremonium coenophialum) -infected tall fescue. Understanding mechanisms responsible for decreased calving and growth rates, delayed onset of puberty, and impaired function of corpora lutea in heifers at puberty consuming endophyte-infected fescue is an emerging field in reproductive toxicology. The condition decreases overall productivity through a reduction in reproductive efficiency, reduced weight gains, and lowered milk production. Reproduction in cattle may be further compromised by winter coat retention, increased susceptibility to high environmental temperatures, and light intolerance. Endocrine effects in steers associated with infected tall fescue include reduced prolactin and melatonin secretions and altered neurotransmitter metabolism in the hypothalamus, the pituitary, and pineal glands. Ewes have decreased prolactin and lengthened intervals from introduction of the ram until conception. The endophyte induces prolonged gestation, thickened placentas, large, weak foals, dystocia, and agalactia in pregnant mares. Ergot peptide alkaloids, produced by the endophyte, are suggested as the primary cause of fescue toxicosis. These compounds reduce prolactin, increase body temperatures, and have powerful vasoconstrictive effects. Neurohormonal imbalances of prolactin and melatonin, with restricted blood flow to internal organs, may be the principal causes of aberrant reproduction, growth, and maturation in livestock consuming endophyte-infected tall fescue.     

Behavior and viability of the premature neonatal foal after induced parturition

Parturition was induced in 2 groups of mares, less than 300 (n = 49) and 300 to 320 days gestation (n = 31), by the administration of prostaglandin F2 alpha or fluprostenol and oxytocin. Foals were categorized into 4 groups according to their ability to adapt in, and survive, the neonatal period. Group A had no demonstrable coordinating reflexes, were weak from birth, and all died within 90 minutes. Group B had some righting reflexes, but had poor coordination and a weak suck reflex. They showed some improvement for about 2 hours, but all died within 9 hours. Group C foals had a good suck reflex and made attempts to stand. After 24 hours, there was a steady deterioration and death occurred within 48 hours. Group D were initially weak, but showed rapid clinical improvement with good adaptation to the environment and survived for at least 7 days. The overall survival rate for the 80 foals born was only 5%. Most group A foals had gestational ages of less than 300 days, but a few (n = 9) were delivered after 300 days and 2 up to 319 days. The youngest survivor was delivered at 318 days and the 3 other survivors were delivered at 320 days. Aspects of the hazards of prematurely induced parturition were considered to be immaturity and stress of parturition.