Optimization of in vitro embryo production and zygote vitrification for the indigenous Vietnamese Ban pig: The effects of different in vitro oocyte maturation systems

The Vietnamese Ban pig is a precious genetic resource that needs to be preserved. In vitro embryo production from in vitro matured (IVM) oocytes is an important tool for the utilization of cryopreserved porcine sperm. The aim of this study was to compare two media for the IVM of Ban pig oocytes. Immature oocytes were subjected to IVM either in a non‐defined (TCM‐199 + pig follicular fluid) or in a defined base medium (POM + epidermal growth factor). At the end of IVM, the oocytes were in vitro fertilized (IVF) with frozen Ban sperm. Ten hours after IVF, the oocytes were either subjected to orcein staining to check fertilization and maturation status or cultured in vitro for 7 days. There was no difference between the two IVM media in terms of percentages of oocyte maturation and blastocyst production. However, the percentage of male pronuclear formation after IVF and the total cell numbers in blastocysts were higher with the defined system. Zygotes obtained by the two IVM systems survived vitrification at similar rates. In conclusion, the two IVM systems were both effective for the production of Ban pig embryos; however, better embryo quality was achieved with the defined one.     

肉牛同期发情效果影响因素的研究

[目的]为了研究同期发情处理方法、繁殖状况、品种、体重等对肉牛同期发情效果的影响,[方法]选择140头不同繁殖状况、品种、体重的肉牛,随机分为3组,分别采用CIDR+第10天肌注PMSG+第11天肌注PGF_2α（埋栓Ⅰ组）、CIDR+第9天肌注PMSG+第11天肌注PGF_2α（埋栓Ⅱ组）、肌注PGF_2α+PMSG(PGF_2α组)的方法进行同期发情处理。[结果]3种不同处理方法相比,PGF_2α组发情率为87.10%,埋栓Ⅰ组、Ⅱ组母牛发情率分别为97.83%、98.41%,显著高于PGF_2α组（P<0.05）;PGF_2α组处理牛受胎率为83.87%,埋栓Ⅰ组、Ⅱ组处理牛受胎率分别为89.13%、84.13%,高于PGF_2α组,但差异不显著（P>0.05）;育成牛与经产牛发情时间相比,埋栓组12～24 h内出现发情症状的经产牛占比为27.27%,育成牛占比为60%,显著高于经产牛（P<0.05）,2...     

Diversity and evolution of major Panax species revealed by scanning the entire chloroplast intergenic spacer sequences

Ginseng is an important medicinal plant, but almost no genomic information is known for it. For the primary step to understand the Panax genome, we inspected the chloroplast genome sequence diversity and used that to infer the evolution of Panax species using them. We inspected a total of 101 intergenic spacers (IGS) covering 44,563 bp (96.8 % of the total IGS) from four Panax species. Diversity was inspected by three steps: gel electrophoresis, high resolution melting (HRM) analysis, and further confirmation by sequencing. Even though low levels of InDel polymorphism were detected by gel electrophoresis, high levels of reproducible polymorphisms were identified by HRM analysis. Sequencing of the HRM-polymorphic spaces and pair-wise multiple sequence alignments revealed up to 247 sequence variations between species. We have identified 62 IGS showing polymorphism between species. Among them, the trnE-trnT, trnT-psbD, ndhF-rpl32, and rpl14-rpl16 spaces are more informative for studying the diversity of Panax relatives. Phylogenetic analysis and molecular evolution studies revealed that P. notoginseng is most diverged from the other Panax species, with a nucleotide substitution rate of 0.0039. P. quinquefolius shows a close relationship with P. ginseng and P. japonicus, with a nucleotide substitution rate of 0.0009. Meanwhile, no sequence variation was detected between P. ginseng and P. japonicus. Calculation of molecular clocks revealed that P. notoginseng diverged more than 1.30 million years ago from the other Panax species and then, P. quinquefolius diverged from P. ginseng by migration in the American continent more than 0.29 million years ago.     

Influence of phosphate sorption on dispersion of a Ferralsol

Abstract

Soil dispersion induces soil erosion and colloidal leaching. Nutrients are lost at the same time and this causes water contamination. Phosphate is an essential element for living organisms. Because phosphate influences soil dispersion and it is an important limited resource, this influence must be evaluated well in order to diminish negative effects on soil structure. In this paper, we firstly evaluated the influence of phosphate sorption on soil dispersion by calculating repulsive potential energy between soil particles. Ferralsol, which is a typical soil in rainy tropical regions, was used as the material. The dispersion-flocculation phenomena were investigated with absorbance of soil suspension under different pH, phosphate adsorption and electrolyte concentration in an Na-NO₃-PO₄ system. The repulsive potential energy was calculated based on the diffuse double layer theory and the measured zeta potential. We indicated that the measured absorbance increased with the increase of the repulsive potential energy. The repulsive potential energy increased with increasing phosphate sorption up to about 5 to 20 mmol kg^?1 at all pH, and it induced the soil dispersion, because phosphate sorption increased the negative charge of the soil. After its peak, it decreased with increasing phosphate sorption because the electrolyte concentration increased and the electrolyte screened the electric field near the soil surface. The repulsive potential energy also increased with increasing pH because of the increase of the negative charge of the soil. Even at low pH, after a certain amount of phosphate sorption, the soil dispersed due to the increase of repulsive potential energy, although the soil flocculated before phosphate application. Because the soil dispersion causes soil and phosphorus loss, the influence of soil pH and phosphate sorption on the soil dispersion should be considered for good soil management.     

测量水技术在宁夏灌区的应用与发展

为实现灌区测量水规范化、标准化和自动化提供参考依据,保证灌区有限水资源科学管理与合理配置,达到公平和高效的利用目的,针对宁夏灌区测量水技术的发展现状进行了综合阐述.总结宁夏目前所采用的量水方法和在生产实际应用中存在的问题,提出灌区量水技术的发展趋势,通过提高灌区测量水的技术装备和测试手段,旨在提高灌区测量水技术水平和精度的有效保证.     

羊痘病毒属病毒通用Cycleave PCR检测方法的建立

为快速特异检测羊痘病毒属病毒（CaPVs）,比较了牛结节性皮肤病病毒（LSDV）、山羊痘病毒（GTPV）和绵羊痘病毒（SSPV）的全基因组序列,选择保守区域设计特异性引物和探针,建立了一种CaPVs通用的Cycleave PCR检测方法。特异性结果显示,该方法在40 min内即可检出CaPVs,与口蹄疫病毒、羊口疮病毒、牛病毒性腹泻病毒、牛传染性鼻气管炎病毒、蓝舌病病毒均无交叉反应;灵敏性结果显示,最低检出限为4.98 copies/μL;重复性结果显示,批内和批间重复变异系数均小于2%。对131份临床样本进行检测发现,该方法与世界动物卫生组织（WOAH）推荐的常规PCR方法相比,敏感性为100%,特异性为91.55%,总符合率为95.42%。以上结果表明,本研究建立的Cycleave PCR检测方法特异、敏感、重复性好,且操作简便,适用于CaPVs的高通量、快速检测。     

Effects of road transportation on metabolic and immunological responses in Holstein heifers

This study examined the effects of road transportation on metabolic and immunological responses in dairy heifers. Twenty Holstein heifers in early pregnancy were divided into non‐transported (NT; n = 7) and transported (T; n = 13) groups. Blood was collected before transportation (BT), immediately after transportation for 100 km (T1) and 200 km (T2), and 24 h after transportation (AT). The T heifers had higher (P < 0.05) blood cortisol and non‐esterified fatty acid concentrations after T1 and T2 than did NT heifers. By contrast, the T heifers had lower (P < 0.05) serum triglyceride concentrations after T1 and T2 than had the NT heifers. The serum cortisol and triglyceride concentrations returned (P > 0.05) to the BT concentrations at 24 h AT in the T heifers. The granulocyte‐to‐lymphocyte ratio and the percentage of monocytes were higher (P < 0.05) after T2 in the T heifers than in the NT heifers, suggesting that transportation stress increased the numbers of innate immune cells. T heifers had higher (P < 0.01) plasma haptoglobin concentrations than NT heifers 24 h AT. In conclusion, transportation increased cortisol secretion and was correlated with increased metabolic responses and up‐regulation of peripheral innate immune cells in dairy heifers.     

An Octopus-Derived Peptide with Antidiuretic Activity in Rats

Discovering new drug candidates with high efficacy and few side effects is a major challenge in new drug development. The two evolutionarily related peptides oxytocin (OXT) and arginine vasopressin (AVP) are known to be associated with a variety of physiological and psychological processes via the association of OXT with three types of AVP receptors. Over decades, many synthetic analogs of these peptides have been designed and tested for therapeutic applications; however, only a few studies of their natural analogs have been performed. In this study, we investigated the bioactivity and usefulness of two natural OXT/AVP analogs that originate from the marine invertebrate Octopus vulgaris, named octopressin (OTP) and cephalotocin (CPT). By measuring the intracellular Ca²⁺ or cyclic AMP increase in each OXT/AVP receptor subtype–overexpressing cell, we found that CPT, but not OTP, acts as a selective agonist of human AVP type 1b and 2 receptors. This behavior is reminiscent of desmopressin, the most widely prescribed antidiuretic drug in the world. Similar to the case for desmopressin, a single intravenous tail injection of CPT into Sprague-Dawley rats reduced urine output and increased urinary osmolality. In conclusion, we suggest that CPT has a significant antidiuretic effect and that CPT might be beneficial for treating urological conditions such as nocturia, enuresis, and diabetes insipidus.     

In vitro effect of pH variations on rumen fermentation, and in vivo effects of buffers in lambs before and after adaptation to high concentrate diets

An in vitro wheat fermentation study (Exp. 1) was conducted to investigate the effect of pH (pH 7, 6, 5 and 4) on rumen lactate and VFA production. In another study (Exp. 2), lambs were fed high concentrate diets containing either no additions (control), 2% sodium bentonite, 2% limestone, 2% NaHCO3 or 10% alfalfa hay. The effect of these diets was determined on ruminal and systemic measurements before and after dietary adaptation. Effect of the control, bentonite, limestone and NaHCO3 treatments on nutrient digestibilities and mineral retentions after adaptation were also determined. Lactate production in the rumen fluid incubated at pH 7 or 6 was negligible. Reducing incubation pH from 6 to 5 resulted in accumulation of both total and L(+)-lactate, but further reduction to pH 4 did not result in an additional increase in lactate production. Variations in incubation pH or time did not affect the ratio of D to L isomers. Lowering the incubation pH to below 6 reduced total VFA production and increased the acetate to propionate ratio. In Exp. 2, the feeding of buffers or alfalfa hay was effective in maintaining a more normal feed intake and ruminal pH, and reduced ruminal lactate. Blood measurements were not affected by dietary treatment. When animals were adapted to the experimental diets, the dietary buffers did not influence rumen and blood measurements. Lambs fed 2% NaHCO3 digested more (P less than .05) organic matter, crude protein, N-free extract and starch, and 2% limestone increased (P less than .05) fiber digestibility. Dietary buffers tended to increase fecal pH and reduce fecal starch Magnesium retention with 2% bentonite, Ca retention with 2% limestone and Na and Mg retention with NaHCO3 were all increased (P less than .05).     

Extracellular cyclic adenosine monophosphate‐dependent protein kinase A autoantibody and C‐reactive protein as serum biomarkers for diagnosis of cancer in dogs

Protein kinase A, a cyclic adenosine monophosphate (AMP)‐dependent enzyme, normally exists within mammalian cells; however, in cancer cells, it can leak out and be found in the serum. Extracellular cyclic AMP‐dependent protein kinase A (ECPKA) has been determined to increase in the serum of cancer‐bearing dogs. However, there have been no reports in the veterinary literature on serum ECPKA autoantibody (ECPKA‐Ab) expression in dogs with cancer. The aim of this study was to evaluate ECPKA‐Ab and C‐reactive protein (CRP) as serum biomarkers for cancer in dogs. ECPKA‐Ab and CRP levels were detected by an enzyme‐linked immunosorbent assay in serum samples from dogs with malignant tumours (n = 167), benign tumours (n = 42), or non‐tumour disease (n = 155) and from healthy control dogs (n = 123). ECPKA‐Ab and CRP levels were significantly higher in the dogs with malignant tumours than in those with benign tumours or non‐tumour diseases, as well as in the healthy controls (P < 0.001, Kruskal‐Wallis test). There was a significant positive correlation between the neoplastic index, which was developed using ECPKA‐Ab and CRP levels, and the presence of cancer in dogs (P < 0.001); the area under the receiver‐operating characteristic curve was estimated to be >0.85 (P < 0.001). In conclusion, ECPKA‐Ab is a potential serum biomarker for a broad spectrum of cancers. Combined measurement of CRP and ECPKA‐Ab levels in serum improves the sensitivity and accuracy of a diagnosis of cancer in dogs.