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.     

(+)-Totarol from Chamaecyparis nootkatensis and activity against Mycobacterium tuberculosis.

The isolation of (+)-totarol as active compound against Mycobacterium tuberculosis is reported from Chamaecyparis nootkatensis outerbark.     

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.     

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.     

Evaluation of two N cycle models for the prediction of N mineralization from grassland soils in the UK

Abstract. The ability of two nitrogen cycle models, of contrasting complexity, to predict N mineralization from a range of grassland soils in the UK, was evaluated. These were NCYCLE, a simple mass balance model of the N cycle in UK grasslands, and CENTURY, a more complex model simulating long-term C, N, P & S dynamics in grassland ecosystems. The models were tested using field measurements of net N mineralization from a range of grassland soils (differing in soil type, history & management practice), obtained over a 2 year period using a soil core incubation technique. This method was considered to measure the total net release of mineral N from the soil organic matter over a specified time, including N which may have been recycled several times. NCYCLE consistently under-estimated mineralization rates at all sites. By contrast, there was some correlation between CENTURY predictions of net N mineralization and field measurements. This may have reflected the different abilities of the two models to simulate N recycling. Neither model, however, was able to predict adequately the effect of cultivation and reseeding on net N mineralization.     

Examination of the equine foot.

The examination of the foot must be based on observations of the hoof, followed by a careful determination of areas of pain. This must be followed by an assessment of the biomechanical forces on the hoof and limb. Finally, imaging gives insight into the nature of the injury and allows the examiner to prognosticate the outcome.     

Association of tumor-associated antigen on the proliferation of bovine leukemia virus-infected lymphoblastoid B-cell lines.

The association of tumor-associated antigen (TAA) on the proliferation of BLV-infected lymphoblastoid B-cell lines (BL2M3 and BL312) was investigated. Flow cytometric analysis of the expression of TAA with monoclonal antibody (mAb) c143 showed high expression of TAA on the surfaces of BL2M3 and BL312 cells. A large amount of TAA was found in the culture supernatant of BL2M3 and BL312 cells as well as in the lysates of BL2M3 and BL312 cells. Culture supernatant but not lysates of BL2M3 and BL312 cells promoted the growth of either BL2M3 cells or BL312 cells. Furthermore, this growth promoting activity in culture supernatants of BL2M3 and BL312 cells was inhibited in a dose-dependent manner when cultured with mAb c143. These results suggested that TAA may be involved in the growth factor-mediated cell growth of bovine B-lymphoblastoid cell lines expressing TAA on their cell surface.