Alfalfa Phytoestrogen Content: Impact of Plant Maturity and Herbage Components

Legumes contain a range of non‐nutritional phytochemicals that may have health‐promoting effects in humans. In this study, we determined the concentrations of four phytoestrogens (coumestrol, apigenin, luteolin and quercetin) in field‐grown alfalfa (Medicago sativa L.). Differences between plants of different stages of maturity, between plant parts, and different canopy segments were assessed. The concentration of individual phytoestrogen in whole herbage varied between 15 and 225 μg g^?1 dry matter (DM) and was strongly affected by stage of maturity. Coumestrol and apigenin concentrations were highest at early vegetative stages, luteolin and quercetin at early vegetative and late flowering stages. All phytoestrogens were found in lowest concentrations at the early flowering stage (average 68 μg g^?1 DM); stage at which alfalfa is usually harvested when used as a forage source for animals. At vegetative stages, apigenin was the predominant phytoestrogen in herbage followed by coumestrol, the reverse being observed upon initiation of flowering; luteolin and quercetin were found at all stages in similarly lower concentrations. Concentrations of luteolin, quercetin and apigenin were 225, 410 and 690 % greater, respectively, in flowers than in leaves or stems; coumestrol concentration was similar between plant parts. In flowers and stems the predominant phytoestrogens were apigenin and quercetin, followed by coumestrol and luteolin. Similar concentrations (average 26 μg g^?1 DM) of each of the four phytoestrogens were found in leaves. Concentrations through the herbage canopy varied and were greatest at >60 cm from the soil surface for apigenin and coumestrol, but greatest at >60 and 0–20 cm for quercetin and at 0–20 cm for luteolin. The results suggest that if alfalfa is to be used as a source of phytoestrogens and is harvested for the production of herbal supplements or nutraceuticals, management will need to be adapted.     

Effect of Oestrous Synchronization with Estradiol 17β and Progesterone on Follicular Wave Dynamics in Dairy Heifers

An experiment was designed to evaluate the effects of estradiol‐17β (E17β) on follicular wave dynamics and ovulatory response in Holstein heifers receiving either a progestogen ear‐implant (Crestar^®; Intervet International b.v. Boxmeer, The Netherlands) or an intravaginal progesterone‐releasing device [controlled internal drug release‐bovine device (Eazibreed, CIDR‐B^®; Bodinco BV, Alkmaar, The Netherlands)]. For comparison, another group of heifers was also synchronized using Crestar plus an injection of estradiol valerate (EV) and norgestomet as recommended by the pharmaceutical company. Twenty 20–22‐month‐old cycling Holstein heifers were allocated to one of the following treatment groups at random stages of the oestrous cycle: (I) simultaneous insertion of Crestar and intramuscular injection of 3 mg norgestomet and 5 mg EV (Crestar 9 + EV 9); (II) simultaneous insertion of Crestar and intramuscular injection of 5 mg E17β (Crestar 9 + E17β 9); (III) insertion of Crestar followed 2 days later by intramuscular injection of 5 mg E17β (Crestar 9 + E17β 7); or (IV) insertion of CIDR‐B device followed 2 days later by intramuscular injection of 5 mg E17β (CIDR 9 + E17β 7). The CIDR‐B or Crestar implants were removed after 9 days and all heifers received 500 μg Cloprostenol (Estrumate^®, Pitman‐Moore Nederland BV, Houten, The Netherlands). Ovarian ultrasonographic examinations were performed once daily during the synchronization period using a B‐mode scanner equipped with a 7.5 MHz linear‐array transrectal transducer. In addition, heifers were scanned every 12 h after implant/device withdrawal until 3 days after ovulation in order to monitor follicular activity, detect ovulation and subsequent early luteal formation. Detection of oestrus was performed every 6 h for 4 days after device/implant removal. Oestrus was observed 24–32 h before ovulation in all heifers. The mean hours interval from treatment withdrawal to ovulation was not significantly different (84.0 ± 16.5, 77.6 ± 4.1, 73.6 ± 4.1 and 64.0 ± 4.4 h for treatments I, II, III and IV, respectively; p > 0.1). However, the variance for heifers treated with EV + norgestomet was significantly larger (Levene’s Test; p < 0.01) than those treated with E17β. All E17β treatments resulted in dominant follicle suppression and a new wave emerged 4.1 days after treatment compared with 6.6 days for the EV + norgestomet treatment (p < 0.05). The time from emergence of the new ovulatory wave to ovulation was longer for the new wave that emerged after E17β treatment (9.2 ± 0.3 days) than after EV + norgestomet treatment (6.9 ± 0.4 days; p < 0.05). The results of this study suggest that the four treatments used were effective in inducing synchronous behavioural oestrus and ovulation. However, a higher degree of oestrus and ovulation synchrony was observed in heifers treated with E17β than in heifers treated with EV + norgestomet. Synchronization treatments with exogenous E17β or EV + norgestomet at the time of progestin device insertion (Crestar or CIDR‐B) or 2 days later in heifers can regulate a different emergence pattern of ovarian follicular development in randomly cyclic heifers. The E17β was effective in inducing follicular suppression and resulted in the consistent emergence of a new follicular wave.     

Trends in mortality ratios among cattle in US feedlots.

OBJECTIVE: To evaluate trends in feedlot cattle mortality ratios over time, by primary body system affected, and by type of animal. DESIGN: Retrospective cohort study. ANIMALS: Approximately 21.8 million cattle entering 121 feedlots in the United States during 1994 through 1999. PROCEDURES: Yearly and monthly mortality ratios were calculated. Numbers of deaths were modeled by use of Poisson regression methods for repeated measures. Relative risks of death over time and by animal type were estimated. RESULTS: Averaged over time, the mortality ratio was 12.6 deaths/1,000 cattle entering the feedlots. The mortality ratio increased from 10.3 deaths/1,000 cattle in 1994 to 14.2 deaths/1,000 cattle in 1999, but this difference was not statistically significant (P = 0.09). Cattle entering the feedlots during 1999 had a significantly increased risk (relative risk, 1.46) of dying of respiratory tract disorders, compared with cattle that entered during 1994, and respiratory tract disorders accounted for 57.1% of all deaths. Dairy cattle had a significantly increased risk of death of any cause, compared with beef steers. Beef heifers had a significantly increased risk of dying of respiratory tract disorders, compared with beef steers. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that although overall yearly mortality ratio did not significantly increase during the study, the risk of death attributable to respiratory tract disorders was increased during most years, compared with risk of death during 1994. The increased rates of fatal respiratory tract disorders may also reflect increased rates of non-fatal respiratory tract disorders, which would be expected to have adverse production effects in surviving animals.     

Cryopreservation of Embryos of Farm Animals

This review contains two parts. The first part is devoted to the significant steps in cryopreservation of mammalian embryos with emphasis on cattle and sheep that serve as models of reference. These steps are: (1) shortening of cooling and warming processes; (2) addition and dilution of cryoprotectant in one step; (3) introduction of plastic straw as a freezing and dilution container; (4) the choice of ethylene glycol as the quite universal cryoprotectant because of its low toxicity and high permeability; (5) vitrification, a cryopreservation method which enable passage from the liquid to the solid state by extreme elevation of viscosity due to high concentration of cryoprotectants and very rapid cooling. There are several vitrification solutions which contain dimethyl sulphoxide, glycerol, ethylene glycol, or a mixture of them, as basic cryoprotectants. The second part considers some factors affecting the efficiency of cryopreservation concerning (i) the origin of embryos and (ii) the stage of development and species. The origin of embryos (in vivo versus in vitro): in vitro embryos show a chilling and freezing sensitivity associated with their lipid content which can be modified by the culture conditions. Both conventional freezing and vitrification have been used and it seems that vitrification is more adapted to in vitro embryos when some modifications of initial protocols are carried out, particularly the rate of cooling. Thus considerable progress has been achieved by using the open pulled straw method of Vajta which enables the use of a minimum volume of freezing medium (0.5 μl) and a very high cooling rate that permits rapid traversal of the damaging temperature zone, corresponding to chilling sensitivity. The stage of development and species: not only are there differences between species at the same stage of development but in the same species all stages of development do not survive equally under the same freezing protocol. In cattle for example, oocytes and early stages of development in vivo or in vitro do not survive whereas compacted morulae and blastocysts survive very well. In the pig hatched blastocysts survive better than the other stages. Horse embryos have special characteristics that pose problems for successful freezing. In conclusion, a lot of work remains to be done to define fundamental characteristics of embryos of certain species (pig, horse) and of embryos of some stages or of oocytes.     

Physeal Fractures in the Horse II. Management and Outcome

Seventy physeal fractures in horses were initially managed by euthanasia (18), stall confinement (25), application of a cast (7), or internal fixation (20). Of the 52 physeal fractures initially managed with stall confinement, a cast, or internal fixation, 23 (44%) healed and 13 (25%) of these horses became sound. The number of horses less than 4.5 months of age with pressure physeal fractures that became sound was significantly higher (p < 0.05) than the number of horses greater than 4.5 months of age. The number of horses with functional, pain-free limbs (sound horses) or functional limbs (lame horses) was not significantly different (p > 0.05) for Salter-Harris Type I, II, III, or IV pressure physeal fractures; however, critical examination for growth disturbances was not performed. The number of horses with pressure or traction physeal fractures of less than 5 days duration prior to presentation that healed or became sound was not significantly different (p > 0.05) when compared with those horses with fractures of greater than or equal to 5 days duration.     

An evaluation of the Olsen test as a measure of plant-available phosphorus in grassland soils derived from basalt parent material

Abstract. Anecdotal and circumstantial evidence have suggested that the Olsen test underestimates plant-available phosphorus (P) in basaltic soils in Northern Ireland. Therefore, the ability of this test to predict plant-available P in basaltic (and non-basaltic) soils was investigated by regressing Olsen-P data against herbage P indices calculated from plant tissue test data using the diagnosis and recommendation integrated system. The average Olsen-P concentration for a range of fields situated on basaltic soils was considerably lower than the average Olsen-P concentration for a range of fields situated on non-basaltic soils, and yet mean sward P status, as given by the herbage P indices, was similar for both groups of fields. Herbage P indices were also much better correlated with Olsen-P measurements in non-basaltic soils than in basaltic soils. Furthermore, at low Olsen-P values (≶9mgPL⁻¹) some swards on basaltic soils were genuinely deficient in P, while others were sufficient or even in surplus for this nutrient. The results confirm that Olsen-P is inadequate as a predictor of plant-available P in basaltic soils. It is concluded that an alternative soil test is needed to provide a reliable assessment of plant-available P in basaltic soils, to prevent overuse of fertilizer and manure P and to minimize the amounts of P entering local watercourses.     

Doxorubicin affects the cardiac muscarinic system in the rat.

During the study on the mechanism of doxorubicin-induced cardiotoxicity, we observed that a long incubation (4 hr) with doxorubicin reduced the maximal negative inotropic effects of a muscarinic receptor agonist, carbachol. The mechanism responsible for this doxorubicin-induced reduction of the efficacy of carbachol was examined in isolated guinea pig hearts. In isolated left atrial muscle preparations, 1 hr incubation with 100 microM doxorubicin caused a parallel right-ward shift of the concentration-response curves for carbachol, but a longer (4 hr) incubation with this agent (30, 100 or 200 microM), caused a significant reduction of the magnitude of the negative inotropic effect of carbachol in addition to the concentration-dependent parallel right-ward shift. The 4-hr incubation with these concentrations of doxorubicin also reduced the maximal negative inotropic effect of an adenosine A1 receptor agonist, R-phenylisopropyl adenosine (R-PIA), without affecting the potency of this agonist. Doxorubicin (1 to 100 microM) reduced [3H]quinuclidinyl benzilate (QNB) binding in a concentration dependent manner, but failed to alter [3HIR-PIA binding. The decrease in the magnitude of the maximal negative inotropic effect by doxorubicin was caused by changes in the muscarinic system at steps common to the transduction of muscarinic and adenosine A1 receptor mechanisms.     

THE EFFECT OF A COMBINATION OF MEDETOMIDINE-BUTORPHANOL AND MEDETOMIDINE, BUTORPHANOL, ATROPINE ON GLOMERULAR FILTRATION RATE IN DOGS

The effects of intramuscularly administered medetomidine and butorphanol (MB), and medetomidine, butorphanol, atropine (MBA) on glomerular filtration rate (GFR) were determined in six dogs as measured by 99m-Tc-labeled diethylenetriaminepentaacetic acid (99mTc-DTPA) nuclear scintigraphy. Direct systolic, diastolic, and mean arterial blood pressures and heart rate were measured at regular time intervals before, during, and after GFR calculations. The mean GFR measurement following MB was significantly greater (4.44 ml/min/kg) than following MBA (3.82 ml/min/kg) or saline treatment (3.41 ml/min/kg). There was no significant difference between the mean GFR measurements following MBA injection and following saline injection. Diastolic and mean arterial pressures following MBA injection were significantly higher than the values recorded after either MB or saline alone. Heart rate following MB administration was significantly lower than that recorded for dogs receiving MBA or saline alone. The results of this study indicate that the administration of medetomidine in combination with butorphanol significantly increases total GFR in healthy dogs, while the administration of the combination of medetomidine, butorphanol, and atropine does not.     

Reproduction in three species of rainbowfish (Melanotaeniidae) from rainforest streams in northern Queensland, Australia

Abstract – The reproductive biology of three species of rainbowfish (Melanotaeniidae) in northeastern Australian rainforest streams was investigated. Two species, Melanotaenia eachamensis and Cairnsichthys rhombosomoides are endemic to the area, whereas the third, M. splendida splendida, is more widespread. The species were all highly fecund, producing many hundreds of eggs between 1.10 and 1.24 mm in diameter. Melanotaenia eachamensis was the most fecund, produced the largest eggs of the three species, and consequently exhibited the greatest maternal investment (as measured by gonadosomatic index). The majority of reproductive effort occurred during the dry season, although reproductively active fish were present year-round for each of the species, but particularly so for M. s. splendida and C. rhombosomoides. No evidence for a role by temperature or photoperiod as environmental cues for reproduction was found, and it was suggested that gonad development was strongly tied to somatic growth. The concentration of reproduction to the dry season ensures that larvae are produced during a period of relatively stable and benign physical conditions. Comparison of temporal changes in gonadosomatic index values suggest that the spawning season of M. eachamensis , which occurs in high-elevation streams, is more restricted and commences about 1 month earlier than either other species. A similar phenology was observed for the M. s. splendida population found at high elevation and highlights the potential for spatial differences in stream productivity to influence life history. ^Note     

Ionic Ratios and Crop Performance: II. Effects of Interactions amongst Vanadium, Phosphorus, Magnesium and Calcium on Soybean Yield

Because vanadium (V) is easily reduced to a cationic form within plant cells, data from resin-extraction of soil were analysed for evidence of interactions between V and the resin-extractable concentrations of magnesium (Mg) and calcium (Ca) on soybean seed yield. Three varieties, 9091, 9061 and 704, were grown over a 3-year period in a corn–soybean–wheat rotation. Surface soil samples (0–15 cm) were extracted with ion-exchange resins, extracts were analysed by inductively coupled plasma methods (ICP), and the results were regressed against seed yield using SAS PROC STEPWISE analysis using forward selection, backward elimination and maximum R² routines. The seed yield of each variety showed a correlation with a unique set of resin-extractable concentrations of V, phosphorus (P), Mg and Ca, and the V:(V + P), Mg:(Mg + Ca), Mg:(Mg + 1000 V) and Ca:(Ca + 1000 V) ratios. Variety 9091 was most sensitive to the Mg:(Mg + Ca) ratio. Variety 9061 was most sensitive to extractable V and to the V:(V + P) ratio. Variety 704 was sensitive to extractable P, V and Ca and the Mg:(Mg + 1000 V) ratio. For variety 9091, Mg fertilization (not currently practised) may be an economical practice, whereas P fertilization of 704 may not be economical. Each regression technique varied slightly in identification of important factors in seed yield. Concentrations and ratios of resin-extractable elements in soil provide insights into optimal genotype selection and possible management alternatives for a given soil.