Effects of FGF10 on Bovine Oocyte Meiosis Progression,Apoptosis, Embryo Development and Relative Abundance of Developmentally Important Genes In Vitro

Fibroblast growth factor (FGF10) acts at the cumulus oocyte complex, increasing the expression of cumulus cell expansion‐related genes and oocyte competency genes. We tested the hypothesis that addition of FGF10 to the maturation medium improves oocyte maturation, decreases the percentage of apoptotic oocytes and increases development to the blastocyst stage while increasing the relative abundance of developmentally important genes (COX2, CDX2 and PLAC8). In all experiments, oocytes were matured for 22 h in TCM‐199 supplemented with 0, 2.5, 10 or 50 ng/ml FGF10. In Experiment 1, after maturation, oocytes were stained with Hoechst to evaluate meiosis progression (metaphase I, intermediary phases and extrusion of the first polar body) and submitted to the TUNEL assay to evaluate apoptosis. In Experiment 2, oocytes were fertilized and cultured to the blastocyst stage. Blastocysts were frozen for analysis of COX2, CDX2 and PLAC8 relative abundance. In Experiment 1, 2.5 ng/ml FGF10 increased (p < 0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50 ng/ml FGF10 (21, 14 and 12%, respectively) and FGF10 decreased the percentage of oocytes that were TUNEL positive in all doses studied. In Experiment 2, there was no difference in the percentage of oocytes becoming blastocysts between treatments and control. Real‐time RT‐PCR showed a tendency of 50 ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10 ng/ml FGF10 to increase CDX2. In conclusion, the addition of FGF10 to the oocyte maturation medium improves oocyte maturation in vitro, decreases the percentage of apoptotic oocytes and tends to increase the relative abundance of developmentally important genes.     

The effect of elemental Zn boluses on concentrations of Zn in serum and faeces of South American camelids

ABSTRACT

Aims: To monitor the effect of using long-acting Zn boluses on the Zn status of a group of South American camelids, using measurements of concentrations of Zn in faeces and serum.

Methods: As part of a facial eczema (FE) prevention programme, 15 camelids were treated with long-acting Zn boluses designed for preventing FE in sheep. Based on bodyweight, 13 alpacas (Vicugna pacos) received two boluses (26.4?g Zn/bolus) and two llamas (Lama glama) received three boluses. In order to monitor Zn status, measurements were made of concentrations of Zn in serum and faeces immediately prior to bolus treatment (Week 0) and 4, 6, 8 and 10 weeks later. Gamma glutamyl transferase (GGT) activity in serum was measured at Weeks 0 and 8.

Results: Two alpacas regurgitated the boluses; in one case the animal was quickly re-treated but this was not possible in the second animal. Mean concentrations of Zn in faeces were higher at all time points compared to Week 0 (p?<?0.001). Peak concentrations were measured at Week 8, and concentrations >120?mg/kg fresh weight (FW), suggested as being protective in calves, were only measured in all (13/13) treated camelids at Week 6. Mean concentrations of Zn in serum differed between weeks of sampling but changes were not consistent, and concentrations did not exceed 18?μmol/L following treatment. There was no evidence of a natural sporidesmin challenge during the study period and activity of GGT in serum of all animals was <45?IU/L.

Conclusions: Treatment with Zn boluses significantly increased concentrations of Zn in faeces but not in serum, but peak concentrations in faeces were only detected 8 weeks after treatment.

Clinical Relevance: The delay in achieving concentrations of Zn in faeces which were associated with protection against FE in calves, combined with the difficulties of administering boluses to camelids, means that we do not believe that Zn boluses should be used as the primary method for preventing FE in camelids. We recommend that FE prevention in camelids should focus on minimising spore production in pasture through the use of fungicides, grazing management and alternative forages, with boluses only used when it is thought that these methods are unlikely to provide sufficient protection against FE. Such use should always be under the guidance of a veterinarian and monitoring of serum GGT activity should be used to ensure that FE control is being achieved.     

Activation of Zona‐Free Buffalo (Bubalus bubalis) Oocytes by Chemical or Electrical stimulation,and Subsequent Parthenogenetic Embryo Development

Parthenogenetic activation using zona‐free oocytes offers an alternative model that could be applied to develop protocols for the activation of reconstructed embryos for cloning. The aim of this study was to compare the efficacy of different methods for the activation of zona‐free buffalo oocytes in terms of their effects on the developmental competence of parthenogenetic embryos. The effects of zona removal on parthenogenetic activation and in vitro developmental competence of metaphase II oocytes were also examined. All activation methods were followed by incubation of 2 mm 6‐dimethylaminopurine (6‐DMAP) for 4 h. Out of three different pulse strengths (1.2, 2.1 or 3.3 kV/cm) used, 2.1 kV/cm resulted in the highest blastocyst rate (25.3%). On comparing different chemical agents and electric pulse, highest blastocyst rate was observed for calcium ionophore (CaI) (28.6%) followed by ethanol (25.0%), electric pulse (22.5%) and combined CaI and ethanol treatment (16.7%) although differences among them were not significant. Furthermore, a significantly reduced developmental potential was observed in zona‐free oocytes when compared to zona‐intact ones up to the blastocyst stage (44.3% vs 27.1%). In conclusion, zona‐free buffalo oocytes can be successfully activated for parthenogenetic development using chemical or electrical stimulation. Out of different agents examined, CaI followed by 6‐DMAP resulted in the highest blastocyst rate.     

Impact of lower atmospheric carbon dioxide on tropical mountain ecosystems

Carbon-isotope values of bulk organic matter from high-altitude lakes on Mount Kenya and Mount Elgon, East Africa, were 10 to 14 per mil higher during glacial times than they are today. Compound-specific isotope analyses of leaf waxes and algal biomarkers show that organisms possessing CO2-concentrating mechanisms, including C4 grasses and freshwater algae, were primarily responsible for this large increase. Carbon limitation due to lower ambient CO2 partial pressures had a significant impact on the distribution of forest on the tropical mountains, in addition to climate. Hence, tree line elevation should not be used to infer palaeotemperatures.     

Resynchronising returns to service in anoestrous dairy cows in the South Island of New Zealand

AIMS: To investigate the effect of targeted resynchronisation of cows treated for non-observed oestrus before the planned start of mating (PSM), that were not detected in oestrus or pregnant 23 days after treatment (phantom cows), on the proportion pregnant at 42 days after PSM and the end of mating.

METHODS: Farm staff from eight herds in two regions of the South Island of New Zealand identified 1,819 cows not showing oestrus by 10 days before PSM. These cows were treated with intravaginal progesterone for 7 days, and I/M gonadorelin 10 days and 1 day before PSM. Three days before PSM they were injected with cloprostenol and equine chorionic gonadotrophin, with fixed time artificial insemination (FTAI) at PSM. By 23 days after PSM, 1,218 cows had not returned to oestrus. Of these, 161 cows confirmed not pregnant by transrectal ultrasonography were randomly assigned to no treatment (control group; n=74) or were resynchronised 25 days after PSM using the same treatment programme as above, with FTAI 35 days after PSM (n=87). All cows that returned to oestrus were artificially inseminated until 42 days after PSM, when natural mating was used. All cows were examined using transrectal ultrasonography 80 to 90 days after PSM to confirm conception dates.

RESULTS: Of the 1,819 anoestrous cows treated before PSM, 526 (29 (95% CI=23.1–34.0)%) had not been observed in oestrus by 23 days after PSM and had not conceived, so were diagnosed as phantoms cows. For resynchronised cows, 42/87 (48 (95% CI=37.8–58.8)%) were pregnant by 42 days after PSM compared to 21/74 (28 (95% CI=18.1–38.7)%) control cows (p=0.009). At the end of mating 58/87 (67 (95% CI=56.6–76.7)%) cows in the resynchronised group were pregnant and 46/74 (62 (95% CI=50.9–73.2)%) in the control group (p=0.554). The hazard of conception from 21 to 42 days after PSM was 1.9 (95% CI=1.07–3.12) times greater for resynchronised than control cows (p=0.026).

CONCLUSION: In cows not observed in oestrus and treated before PSM, resynchronisation increased the proportion pregnant by 42 days after PSM.

CLINICAL RELEVANCE: The benefit of resynchronisation depends on the number of anoestrous cows before PSM and the number of phantom cows after PSM. However at the herd-level it is likely that providing advice to reduce the known risk factors for cows not being observed in oestrus before the PSM may well be more cost effective than identifying and treating a sub-population of phantom cows.     

Osmium Recycling in Subduction Zones

Peridotite xenoliths from the Cascade arc in the United States and in the Japan arc have neodymium and osmium isotopic compositions that are consistent with addition of 5 to 15 percent of subducted material to the present-day depleted mantle. These observations suggest that osmium can be partitioned into oxidized and chlorine-rich slab-derived fluids or melts. These results place new constraints on the behavior of osmium (and possibly other platinum group elements) during subduction of oceanic crust by showing that osmium can be transported into the mantle wedge.     

A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand

ABSTRACT

The aim of this review is to critically assess the test characteristics and practicality of published data on direct and indirect tests for diagnosing failure of transfer of passive immunity (FPT) in dairy calves in New Zealand, to provide recommendations for veterinary practitioners, and to examine the recommended sample size for assessing herd-level prevalence of FPT and the confidence in the results obtained. The definition of FPT is based on measurement of concentrations of IgG in serum of neonatal calves after colostrum intake. The gold standard method for measurement of concentrations of IgG is radial immunodiffusion. However its cost, requirements for laboratory equipment, and the time taken to obtain results have meant that alternative tests have been developed. The turbidimetric immunoassay and ELISA also directly measure concentrations of IgG. Indirect tests include measurement of concentrations of total proteins (TP) in the laboratory or using a refractometer, γ-glutamyl transferase (GGT) activity, and the zinc sulfate turbidity (ZST) test. Of the indirect tests, measurement of concentrations of TP in the laboratory or using a refractometer combine high specificity and sensitivity with a consistent association with concentrations of IgG in calves between 1–7 days of age. Using a refractometer is less accurate than direct measurement in a laboratory, but is still a suitable test if low cost and speed are important. Although GGT activity is strongly associated with concentrations of IgG in serum, the relationship varies with time after birth. Therefore the target thresholds change with time, increasing error compared to the measurement of concentrations of TP in serum. Similarly, factors other than total concentrations of IgG have a significant effect on the association with ZST test, complicating interpretation. Thus, when direct measurement of concentrations of IgG is not feasible, the recommendation is that concentrations of TP in serum should be used as the diagnostic test for diagnosis of FPT, providing calves are not dehydrated. Using a sample size of 12 calves is suitable for estimating whether the herd-level prevalence of FPT is <20% or >20%, if there are no calves or >5 calves diagnosed with FPT, respectively, but is limited in diagnostic confidence when 1–4 calves test positive. Diagnostic interpretation can be significantly improved if tests of FPT are used alongside information on the likely risk of FPT on the tested farm.     

黄土的红外反射光谱与红外光声光谱特征及其差异研究

红外光谱法作为一种新的研究手段已经广泛应用于土壤分析,由其检测区域和手段的不同又可分为多种光谱类 型。本研究以第四纪黄土为例,系统地比较了近红外区和中红外区反射光谱和光声光谱的吸收特征及其差异。结果表明,中红外光谱比近红外光谱的信息更为丰富,且中红外光谱与样品中物质的特征吸收关系更加密切,从而更有利于土壤定性与定量分析。土壤的反射光谱和光声光谱表现出了明显不同的特征,在近红外区,反射光谱和光声光谱吸收明显不同,而在中红外区,反射光谱和光声光谱具有相对应的吸收,但相对吸收强度明显不同,且吸收峰的位置也发生改变,尤其在1 000 ~ 2 000 cm-1谱区,反射光谱相互干扰很强,而光声光谱的吸收特征更为明显。在黄土的分类鉴别上,反射光谱优于光声光谱。红外反射光谱和光声光谱在不同波段下具有不同的吸收灵敏度,在土壤定性与定量分析中各自都将具有其明显的优势。