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.     

Direct and Residual Contributions of Symbiotic Nitrogen Fixation by Legumes to the Yield and Nitrogen Uptake of Maize (Zea mays L.) in the Nigerian Savannah

Field experiments were conducted to determine the direct and residual contributions of legumes to the yield and nitrogen (N) uptake of maize during the wet seasons of 1994 and 1995 at the University Farm, Abubakar Tafawa Balewa University, Bauchi, Nigeria, located in the Northern Guinea savannah of Nigeria. Nodulating soybean, lablab, green gram and black gram contributed to the yield and N uptake of maize either intercropped with the legumes or grown after legumes as a sole crop. Direct transfer of N from the nodulating soybean, lablab, green gram and black gram to the intercropped maize was 24.9–28.1, 23.8–29.2, 19.7–22.1 and 18.4–18.6 kg N ha^–1, respectively. However, the transfer of residual N from these legumes to the succeeding maize crop was 18.4–20.0, 19.5–29.9, 12.0–13.7 and 9.3–10.3 kg N ha^–1, respectively. Four years of continuous lablab cropping resulted in yields and N uptake of the succeeding maize crop grown without fertilizer N that were comparable to the yields and N uptake of the succeeding maize crop supplied with 40–45 kg N ha^–1 and grown after 4 years of continuous sorghum cropping. It may therefore be concluded that nodulating soybean, lablab, green gram and black gram may be either intercropped or grown in rotation with cereals in order to economize the use of fertilizer N for maize production in the Nigerian savannah.     

The Morphology of 16 and 17 Day Old Bovine Blastocysts from Virgin and Repeat Breeder Heifers

The morphology of 16–17 days old embryos from virgin heifers (VH) and repeat breeder heifers (RBH) was compared using light and electron microscopy. In addition some embryos transferred from one heifer category to the other were studied. Embryos from VH were elongated blastocysts and the oval embryonic disc had three germ layers. The ectoderm was stratified and many mitoses were seen. The endoderm lining the blastocoelic cavity consisted of almost squamous cells conjoined by tight junctions. Between the ectoderm and the endoderm the mesoderm had developed and expanded laterally and the coelom had formed. The trophoblastic cells adjacent do the embryonic disc were cylindrical, whereas those more peripheral located were cuboidal. The trophoblastic cells were conjoined by tight junctions and they had numerous long microvilli on their peripheral surface. Except in the embryonic disc region, the endodermal cells had filopodial processes towards the trophoblast. The embryos from RBH varied in appearance. One was similar to those from VH whereas the others were, more or less retarded, without formation of mesoderm. The smaller one consisted ot trophoblastic cells only. The transferred embryos (representing surviving embryos: 2 out of 9 in VH-RBH and 5 of 6 in RBH-VH) had a morphology similar to that of VH blastocytes two though, appeared somewhat retarded. It is suggested that the retarded embryos lack the ability to complete embryonic development and that the uterine environment of RBH is not favourable to sustain normal embryonic development.     

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.     

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     

Validation of the otoliths for age determination in Barbus sclateri (Günth.)

Abstract – Otoliths were used to explore the timing of annulus formation in Barbus sclateri. Lapillus exhibited opaque and translucent zones whose combination was formed once a year. Opaque zones appeared formed from July and January, and the translucent ones from February to June. We describe a procedure to assess age by using the number of annulii, the condition of the otolith's edge and the capture date. The use of otoliths for age determination in this cyprinid endemic to the Iberian Peninsula provides a useful alternative to the traditional methods of scale reading. ^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.     

Acupuncture for general veterinary practice.

Since the International Society of Veterinary Acupuncture (IVAS) was founded in 1974, acupuncture (AP) has received greater acceptance by veterinary professionals throughout the world. This article introduces some important animal diseases that respond well to AP therapy. These include resuscitation of small animals, treatment of anoestrous gilts and sows, bovine reproductive disease, canine vertebral problems and equine backpain, etc. Conventional medicine considers these to be difficult cases to treat. Veterinarians have become more aware of the benefits of AP especially for those diseases, thanks to the efforts of experienced practitioners and scientists, and the many published reports on veterinary AP that have introduced some good indications for AP therapy in veterinary practice. Possible mechanisms behind the effectiveness of AP are discussed. This article aims to introduce veterinarians to good indications for AP to initiate their interest in the practice of AP. Although this is a rapidly expanding field, a long march must begin with one step. We wish this article to be the shoes for such a march. For more information on veterinary AP, contact IVAS , P.O. Box 1478, Longmont, CO.. 80502, USA (http://www.ivas.org), or your national veterinary acupuncture society (http://www.komvet.at/ ivadkom/vapsocs.htm).