Reduced Immunolabelling of a Porcine Oocyte Membrane Protein Reflects Reduced Fertilizability of Porcine Oocytes Following Elevated Ambient Temperature

Decreased fertility in pigs is a common occurrence during summer months. An objective of the current experiments was to evaluate if elevated ambient temperature altered the oocyte plasma membrane including potential receptors for sperm. This would potentially contribute to reduced fertilizability. Treated gilts were exposed in vivo to 32°C for 12 h per day and 20°C for the remaining 12 h per day for 7 days; control gilts were exposed to 22°C for 12 h and 20°C for the remaining 12 h each day. Cumulus–oocyte complexes were also aspirated from ovaries obtained from gilts maintained at thermoneutral ambient temperature and matured in vitro at 38.5°C or 40°C. Relative abundance of a porcine oocyte membrane protein was examined by intensity of immunolabelling of the in vivo and in vitro matured oocytes evaluated with confocal microscopy; fertilizability of the in vitro matured oocytes was evaluated in in vitro fertilization assays. Oocytes obtained from gilts exposed to elevated ambient temperature for 7 days had reduced immunolabelling compared with oocytes from control gilts (p < 0.05). Similarly, oocytes matured in vitro for 44 h at elevated ambient temperature had reduced immunolabelling and reduced fertilizability compared with oocytes matured at 38.5°C (p < 0.01 and p < 0.05). These results suggest porcine oocyte quality is reduced by elevated ambient temperature and immunolabelling of oocytes with antibodies to specific membrane proteins may be effective to evaluate some aspects of oocyte quality.     

Effect of Taurine and Melatonin in the Culture Medium on Buffalo In Vitro Embryo Development

This study was carried out to investigate the effect of supplementing culture medium with different concentrations of taurine and melatonin, on buffalo oocyte in vitro meiotic maturation and embryo development. In experiment 1, oocytes were matured in vitro and the cleaved embryos were cultured in the same following seven culture medium; (i) control (TCM 199 + 10% SS); (ii) control + 0.5 m m taurine; (iii) control + 1 m m taurine; (iv) control + 3 m m taurine; (v) control + 5 μ m melatonin; (vi) control + 10 μ m melatonin and (vii) control + 50 μ m melatonin. In experiment 2, based on the results of experiment 1, to examine the synergistic effect of antioxidants, the oocytes were matured in culture medium (TCM199 + 10% SS), supplemented with both taurine at 1 m m and melatonin at 10 μ m concentration and the cleaved embryos were cultured in the same medium. Supplementation of taurine at 1 m m concentration in the culture medium resulted in a higher (p < 0.05) transferable embryo (TE) yield when compared with control (20.6% vs 14.1%). Supplementation of melatonin at 10 and 50 μ m concentration in the culture medium resulted in a higher (p < 0.05) meiotic maturation rate (90.3% and 88.8% respectively) and TE yield (28.4% and 27.2% respectively), than the other treatments. In experiment 2, the TE yield did not improve by supplementing the culture medium with both taurine and melatonin, when compared with melatonin alone. In conclusion, the results of this study demonstrated that, enriching the culture medium with taurine and melatonin, improves in vitro embryo production efficiency in buffaloes. In particular, a high TE yield was obtained by enriching the culture medium with 10 μ m melatonin.     

Methyl jasmonate treatment of mature Norway spruce (Picea abies) trees increases the accumulation of terpenoid resin components and protects against infection by Ceratocystis polonica, a bark beetle-associated fungus

When conifers such as Picea abies Karst. (Norway spruce) are attacked by insects or pathogens, they often respond by producing increased quantities of terpenoid oleoresin. This response can be mimicked in young P. abies seedlings by treatment with methyl jasmonate (MJ). In this study, we determined the effects of MJ on terpenoids and other chemical defenses of mature P. abies, and investigated whether this treatment protected trees against attack by the blue-stain fungus Ceratocystis polonica (Siem.) C. Moreau, the most important fungal associate of the spruce bark beetle Ips typographus L. Methyl jasmonate treatment induced the formation of traumatic resin ducts in the developing xylem, enhanced resin flow and stimulated increased accumulation of monoterpenes, sesquiterpenes and diterpene resin acids. However, only minor changes were detected in terpene composition in response to MJ treatment and no changes in soluble phenolic concentration were measured. There was much variability in the timing and degree of response to MJ among clones. The observed chemical and anatomical changes in response to MJ treatment were correlated with increased resistance to C. polonica, suggesting that terpenoid oleoresin may function in defense against this pathogen.     

On the mode of action of the herbicides cinmethylin and 5-benzyloxymethyl-1, 2-isoxazolines: putative inhibitors of plant tyrosine aminotransferase

BACKGROUND: The mode of action of the grass herbicides cinmethylin and 5‐benzyloxymethyl‐1,2‐isoxazolines substituted with methylthiophene (methiozolin) or pyridine (ISO1, ISO2) was investigated. RESULTS: Physiological profiling using a series of biotests and metabolic profiling in treated duckweed (Lemna paucicostata L.) suggested a common mode of action for the herbicides. Symptoms of growth inhibition and photobleaching of new fronds in Lemna were accompanied with metabolite changes indicating an upregulation of shikimate and tyrosine metabolism, paralleled by decreased plastoquinone and carotenoid synthesis. Supplying Lemna with 10 µM of 4‐hydroxyphenylpyruvate (4‐HPP) reversed phytotoxic effects of cinmethylin and isoxazolines to a great extent, whereas the addition of L ‐tyrosine was ineffective. It was hypothesised that the herbicides block the conversion of tyrosine to 4‐HPP, catalysed by tyrosine aminotransferase (TAT), in the prenylquinone pathway which provides plastoquinone, a cofactor of phytoene desaturase in carotenoid synthesis. Accordingly, enhanced resistance to ISO1 treatment was observed in Arabidopsis thaliana L. mutants, which overexpress the yeast prephenate dehydrogenase in plastids as a TAT bypass. In addition, the herbicides were able to inhibit TAT7 activity in vitro for the recombinant enzyme of A. thaliana. CONCLUSION: The results suggest that TAT7 or another TAT isoenzyme is the putative target of the herbicides. Copyright © 2011 Society of Chemical Industry     

Kepler-36: a pair of planets with neighboring orbits and dissimilar densities

In the solar system, the planets' compositions vary with orbital distance, with rocky planets in close orbits and lower-density gas giants in wider orbits. The detection of close-in giant planets around other stars was the first clue that this pattern is not universal and that planets' orbits can change substantially after their formation. Here, we report another violation of the orbit-composition pattern: two planets orbiting the same star with orbital distances differing by only 10% and densities differing by a factor of 8. One planet is likely a rocky "super-Earth," whereas the other is more akin to Neptune. These planets are 20 times more closely spaced and have a larger density contrast than any adjacent pair of planets in the solar system.     

Variability in Namibian landraces of watermelon (Citrullus lanatus)

An on-station experiment was carried out in northern Namibia to compare local landraces of watermelon to modern varieties. Three groups of local watermelon are distinguished by farmers in Namibia: watermelons for fresh consumption, cooking melons used for porridge, and seed melons for oil pressing. The present study was carried out to estimate agronomic potential of the local landraces and verify whether agronomic characteristics would justify the local groupings. Important agronomic and fruit-quality traits (yield, earliness, fruit weight, fruit number, rind thickness, soluble solids, seed weight, and tolerance to biotic and abiotic stress) were measured in seven landraces (three watermelons, three cooking melons and one seed melon)and compared to three modern cultivars frequently grown in Namibia. The study revealed that well-adapted, productive Citrullus lanatuslandraces are available, and cultivated by local farmers throughout the northern regions of Namibia. Based on a combination of agronomic characteristics these local varieties could be grouped into three distinct types: watermelons, cooking melons, and seed melons, in agreement with farmers classification. Landrace water melons were characterized by smaller, less sweet fruits with larger seed and a thicker rind compared to modern varieties. Yield level was comparable but development was later and fruit number in some cases higher. Cooking water melons were less sweet than local water melon with large fruits comparable in size to modern cultivars. The only included seed watermelon had small fruits with large seeds and low soluble sugars. Yield varied from 118 t/ha for a giant cooking melon, out yielding all other accessions, to only 10t/ha for the seed melon type, but with considerable variation within groups. The earliness of commercial varieties maybe of interest in developing a mixed portfolio of short- and long-cycle landraces, whereas plant vigour, resistance and seed characters in some of the Namibian landraces may be important traits for breeding for both local purposes and for commercial varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of drought stress on content and composition of seed alkaloids in narrow-leafed lupin, Lupinus angustifolius L.

The effect of drought stress at three growth stages on seed alkaloid content in two sweet (W26 and Polonez) and one bitter variety (Zubr) of spring sown Lupinus angustifolius L. was investigated in a lysimeter experiment on sandy soil in Denmark. Full irrigation resulted in medium alkaloid content. Drought stress during the vegetative phase increased alkaloid content to 0.094, 0.192 and 2.55% of seed dry matter, respectively. At the flowering stage alkaloid content decreased to 0.070, 0.152 and 1.99%, while drought stress during pod-filling increased alkaloid content for the determinate variety W26 to 0.097%, but decreased it to 0.161 and 1.98% for the two indeterminate varieties Polonez and Zubr, respectively. Drought during either flowering or pod-filling resulted in an increased mean seed weight and a reduced number of seed per square metre. Alkaloid content was negatively correlated to seed weight. The alkaloid composition differed for the three genotypes, and was affected by drought treatment. Compared to the bitter variety the sweet varieties were characterized by reduced concentrations of lupanin, 13-hydroxylupanin and angustifolin, whereas spartein, isolupanin and isoangustifolin were little affected.     

The Predictive Value of Semen Analysis in the Evaluation of Stallion Fertility

Pregnancy rates in managed horse populations depend on the innate fertility of the mares and stallions involved and on the quality of breeding management. Of course, because a single stallion usually mates many mares, stallion fertility is a critical factor in the overall success of a breeding program. Unfortunately, accurate evaluation of stallion fertility per se requires a large number of normal mares to be mated and is necessarily retrospective. Rather, the ideal is to predict fertility in advance of the stallion's breeding career, and this is currently attempted by way of a thorough physical examination and a routine analysis of semen quality. However, while such a ‘breeding soundness examination’ identifies stallions that clearly lack the capacity for adequate fertility, it is of limited use for predicting the level of fertility and fails to identify some seriously sub‐fertile animals. Similarly, while various sperm function tests (e.g., sperm head morphometry, the hypoosmotic swelling test, glass wool‐sephadex filtration, progesterone receptor exposure) have been shown to correlate fairly well with fertility in the field, most examine only a single or a narrow range of the attributes that a sperm must possess if it is to fertilize an oocyte in vivo, and are thus more useful for identifying specific causes of sub‐fertility than for predicting the level of fertility. On the other hand, combining the results of the various sperm function tests does improve the reliability of fertility estimation and current research is therefore concentrated on identifying a range of tests that covers as many important sperm attributes as possible but that can be performed rapidly and cheaply. In this respect, flow‐cytometry has proven to be an ideal tool because it allows the objective, rapid and simultaneous analysis of a number of properties in a large number of sperm. Moreover, stains are available for an increasing range of sperm characteristics including viability, capacitation and acrosome status, mitochondrial activity and chromatin integrity. Flow‐cytometric analysis of sperm with appropriate probes thus offers considerable promise for the prediction of stallion fertility.     

Nitrogen utilization and digestibility of amino acids by lambs fed a high-concentrate diet with limestone or magnesium oxide

Effects were evaluated of high dietary levels of magnesium oxide (MgO) or limestone on DM, OM and CP digestibility, N balance and intestinal absorption of amino acids by lambs fed a high concentrate diet. Twelve wether lambs equipped with abomasal and ileal cannulas were blocked by weight and breeding and allotted to treatments in a randomized block design in two consecutive trials. Diets were control (800 g), control plus 1.5% MgO (812 g), control plus 1.5% limestone (812 g) and control plus 3.0% limestone (824 g) fed in two equal portions at 12-h intervals. Ruminal fluid pH differences were small. The pH of ileal digesta was greater (P less than .05) with MgO than with limestone (8.23 vs 7.73). Fecal pH was higher (P less than .01) for lambs fed all mineral treatments (avg 8.75) than for lambs fed the control (7.61) and was higher (P less than .01) when MgO (9.53) rather than limestone (8.36) was fed. Ruminal NH3N was lower (P less than .01) when lambs were fed MgO (11.9 mg/dl) compared with limestone (avg 31.2 mg/dl). Preintestinal DM digestibility was greatest (P less than .10) with limestone (avg 49.5%) feeding compared with feeding MgO (31.2%) or the control (35.4%). About 41.5% more essential (P less than .05) and 48% more nonessential (P less than .03) amino acids reached the small intestine when MgO was fed than when limestone was fed. Partial digestibility of amino acids in the small intestine was reduced (P less than .03) an average of 5 percentage units when MgO or limestone was fed. Feeding high levels of MgO or limestone to lambs did not improve the overall digestibility of DM, OM or CP. In fact, feeding high levels of MgO or limestone appeared to be detrimental, reducing intestinal absorption of amino acids.     

Horse species symposium: a novel approach to monitoring pathogen progression during uterine and placental infection in the mare using bioluminescence imaging technology and lux-modified bacteria

Uterine and placental infections are the leading cause of abortion, stillbirth, and preterm delivery in the mare. Whereas uterine and placental infections in women have been studied extensively, a comprehensive examination of the pathogenic processes leading to this unsatisfactory pregnancy outcome in the mare has yet to be completed. Most information in the literature relating to late-term pregnancy loss in mares is based on retrospective studies of clinical cases submitted for necropsy. Here we report the development and application of a novel approach, whereby transgenically modified bacteria transformed with lux genes of Xenorhabdus luminescens or Photorhabdus luminescens origin and biophotonic imaging are utilized to better understand pathogen-induced preterm birth in late-term pregnant mares. This technology uses highly sensitive bioluminescence imaging camera systems to localize and monitor pathogen progression during tissue invasion by measuring the bioluminescent signatures emitted by the lux-modified pathogens. This method has an important advantage in that it allows for the potential tracking of pathogens in vivo in real time and over time, which was hitherto impossible. Although the application of this technology in domestic animals is in its infancy, investigators were successful in identifying the fetal lungs, sinuses, nares, urinary, and gastrointestinal systems as primary tissues for pathogen invasion after experimental infection of pregnant mares with lux-modified Escherichia coli. It is important that pathogens were not detected in other vital organs, such as the liver, brain, and cardiac system. Such precision in localizing sites of pathogen invasion provides potential application for this novel approach in the development of more targeted therapeutic interventions for pathogen-related diseases in the equine and other domestic species.