Formation of aerobic granules and their PHB production at various substrate and ammonium concentrations

Aerobic granular sludge rich in polyhydroxybutyrate (PHB) was cultivated in a sequencing batch reactor (SBR) by seeding anaerobic granular sludge. The PHB content in aerobic granules was investigated and the experimental results reveal that both influent chemical oxygen demand (COD) and ammonium concentrations had a significant effect on the morphological characteristics and the PHB production of the aerobic granular sludge. At a COD and ammonium concentration of 750 mg/L and 8.5mg/L, respectively, the PHB content of the granules reached 44%, but their poor settling ability, as evidenced by a high sludge volume index, was observed. This was attributed to the outgrowth of filamentous bacteria on the granule surface. However, an increase in the ammonium concentration resulted in an elevated sludge concentration and a decrease in the PHB content in the granules. In this case, the aerobic granular sludge with a regular and compact structure was formed. The results suggest that, through controlling the COD and ammonium concentrations in the influent, the PHB-rich aerobic granular sludge with good settling ability could be cultivated.     

原核生物蛋白O-糖基化的研究进展

自从在原核生物中发现蛋白糖基化之后,越来越多的O-糖基化机制在不同种属的细菌中被发现。本文根据对O-寡糖基转移酶(O-oligosaccharide transferase,OTase)的依赖与否,将原核生物的O-糖基化分为OTase非依赖型和OTase依赖型,并分别对这两种糖基化机制进行了详细阐述。通过对不同的O-糖基化机制的深入了解,为以后更好地利用这些途径来合成工程化的目标糖蛋白奠定基础。     

Generation of buffalo mammary epithelial cells with targeted knockout of 18s rDNA by a CRISPR/Cas9 adenovirus system

In recent years, CRISPR/Cas9 has rapidly become one of the most promising genome editing tools because it is simple and easy to use and cost effective. However, the large size of Cas9 sequences limits its application in clinically promising vectors and it also impacts non-viral transfection. In this study, CRISPR/Cas9 adenovirus vectors that target the buffalo 18s rDNA gene were constructed, transfected into 293 cells for adenovirus packaging, and the adenovirus was used to knockout the 18s rDNA gene in buffalo mammary epithelial cells. The results demonstrated that the CRISPR/Cas9 adenovirus vectors for the buffalo 18s rDNA gene could efficiently target the sites as revealed by the fluorescence reporter system. After amplification, the adenovirus titer of Sn458-18s1 and Sn458-18s2 reached 1.03 × 10⁹PFU/mL and 1.05 × 10⁹ PFU/mL, respectively. For buffalo mammary epithelial cell infection, the efficiency was 100% when the multiplicity of infection (MOI) ≥ 100 PFU/mL. There were 9 mutational clones found in the 20 clones, and the gene mutagenesis rate reached 45%. Of these, 2 clones were 35-bp deleted and 7 clones were 12-bp deleted. These results suggested that the adenovirus system overcame the low transfection efficiency of the buffalo mammary epithelial cells associated with using lipid-based methods or electroporation. Moreover, we preliminary developed an efficient technique for multiple-locus gene targeting at repeated sequences of the buffalo genome.     

大兴安岭泥炭地植物叶片碳氮磷含量及其化学计量学特征

叶片碳(C)、氮(N)、磷(P)含量及其化学计量特征为植物养分状况和元素限制性提供依据。为了解不同生活型植物叶片C、N、P化学计量特征的变化,该研究测定、分析了大兴安岭地区18个泥炭地常见的3种草本植物——白毛羊胡子草(Eriophorum vaginatum)、玉簪薹草(Carex globularis)、小叶章(Deyeuxia angustifolia), 5种落叶灌木——柴桦(Betula fruticosa)、越桔柳(Salix myrtilloides)、细叶沼柳(Salix rosmarinifolia)、笃斯越桔(Vaccinium uliginosum)、越桔(Vaccinium vitis-idaea)和3种常绿灌木——杜香(Ledum palustre)、地桂(Chamaedaphne calyculata)、头花杜鹃(Rhododendron capitatum)的叶片C、N、P含量。结果表明: (1)落叶和常绿灌木叶片C、N、P含量总体高于草本植物而C:N、C:P、N:P低于草本植物, 说明不同生活型植物具有不同的养分利用策略,灌木叶片C、N、P储存高于草本植物而N、P利用效率低于草本植物; (2)小叶章和头花杜鹃叶片N:P小于10, 同时其N含量小于全球植物叶片平均N含量, 相比其他植物来说更易受N限制; (3)采样地点解释了叶片C、N、P指标变异的12.8%-40.8%, 植物种类对叶片C、N、P指标变异的解释量占9.3%-25.5%; (4)草本植物C、N、P指标的地点间变异系数高于落叶和常绿灌木, 草本植物C、N、P指标对地点因素变化的响应较灌木敏感; (5)草本植物N含量种间变异系数高于落叶和常绿灌木, 落叶灌木P含量种间变异系数高于草本植物和常绿灌木, 草本植物和落叶灌木N、P吸收的种间生理分化较常绿灌木高。     

Bioflocculant production by culture of Serratia ficaria and its application in wastewater treatment

A bioflocculant-producing bacterium was isolated from soil and identified as Serratia ficaria. Using optimized culture conditions a flocculating activity of 95.4% was obtained. It was found to be effective for flocculation of a kaolin suspension over weakly acidic pH (5-7); divalent cations (Ca2+ and Mg2+) enhanced the flocculating activity, while the co-presence of Al3+ and Fe3+ resulted the negative effect. Measurements of zeta potential revealed that charge neutralization played an important role in the flocculation. It could flocculate a variety of real wastewaters, including river water, brewery wastewater, meat processing wastewater and soy sauce brewing wastewater. The bioflocculant was also used to treat pulp effluent, and the removal rate of color and chemical oxygen demand (COD) were up to 99.9% and 72.1%, respectively, which were better than traditional chemical flocculants.     

Mitochondrial Genetic Variation in Chinese Pigs and Wild Boars

The mitochondrial DNAs (mtDNAs) from 30 pig breeds (29 Chinese native breeds and 1 European breed) and wild boars were investigated for restriction fragment length polymorphisms (RFLPs) to determine the phylogenetic relationships and genetic diversity among pig breeds and wild boars. Of the 24 enzymes used, 8 (AvaI, BclI, BglII, EcoRI, EcoRV, ScaI, StuI, and XbaI) detected polymorphisms. By combining the cleavage patterns for each enzyme, 108 individuals were sorted into eight mtDNA mitotypes. There are two haplotype lineages in domestic pigs, i.e., Chinese and European lineages. The pairwise nucleotide sequence divergence was calculated to be 0.56% between Chinese pigs and European pigs, suggesting that they might have diverged from a common ancestor approximately 280,000 years ago. The wild boars showed more extensive genetic variation, four mitotypes were detected in six wild boars. In addition, one of the Zhejiang wild boars was found to share the same mitotype with Chinese native pigs. A UPGMA tree based on genetic distance among mitotypes indicated that mtDNAs of Chinese pigs and European pigs are clearly divided into two clusters, and Chinese wild boars are more closely related to the Chinese pigs. Our results provide molecular evidence to support the previous hypothesis that pigs may be derived from two maternal origins, Asian and European wild boars. Chinese native pig breeds may have a single origin.     

Use of an Autologous Liver Round Ligament Flap Zeros Postoperative Bile Leak after Curative Resection of Hilar Cholangiocarcinoma

BackgroundPostoperative bile leak is a major surgical morbidity after curative resection with hepaticojejunostomy for hilar cholangiocarcinoma, especially in Bismuth-Corlette types III and IV. This retrospective study assessed the effectiveness and safety of an autologous hepatic round ligament flap (AHRLF) for reducing bile leak after hilar hepaticojejunostomy.MethodsNine type III and IV hilar cholangiocarcinoma patients were consecutively hospitalized for elective perihilar partial hepatectomy with hilar hepaticojejunostomy using an AHRLF between October 2009 and September 2013. The AHRLF was harvested to reinforce the perihilar hepaticojejunostomy. Main outcome measures included operative time, blood loss, postoperative recovery times, morbidity, bile leak, R0 resection rate, and overall survival.ResultsAll patients underwent uneventful R0 resection with hilar hepaticojejunostomy. No patient experienced postoperative bile leak.ConclusionsThe AHRLF was associated with lack of bile leak after curative perihilar hepatectomy with hepaticojejunostomy for hilar cholangiocarcinoma, without compromising oncologic safety, and is recommended in selected patients.     

Genetic Diversity and Relationship of Yunnan Native Cattle Breeds and Introduced Beef Cattle Breeds

In this study, random amplified polymorphic DNA (RAPD) analysis was used to estimate genetic diversity and relationship in 134 samples belonging to two native cattle breeds from the Yunnan province of China (DeHong cattle and DiQing cattle) and four introduced beef cattle breeds (Brahman, Simmental, MurryGrey, and ShortHorn). Ten primers were used, and a total of 84 bands were scored, of which 63 bands (75.0%) were polymorphic. The genetic distance matrix was obtained by proportions of shared fragment. The results indicate that the Yunnnan DeHong cattle breed is closely related to the Brahman (Bos indicus), and the Yunnan DiQing cattle breed is closely related to the Simmental, ShortHorn, and MurryGrey (Bos taurus) breeds. Our results imply that Bos indicus and Bos taurus were the two main origins of Yunnan native cattle. The results also provide the basic genetic materials for conservation of cattle resources and crossbreeding of beef cattle breeds in South China.