Isolation and characterization of 23 microsatellite loci in the Chinese sucker (Myxocyprinus asiaticus)

Chinese sucker (Myxocyprinus asiaticus) is a second class state-protected animal in China. In this study, we developed twenty-three polymorphic microsatellite loci in Chinese sucker. The observed and expected heterozygosity ranged from 0.292–0.958 to 0.423–0.900, respectively. The polymorphic information content ranged from 0.356 to 0.869, with a mean of 0.710. These microsatellite loci are expected to be useful for further studies of genetic diversity, population genetic structure, and assessments of the artificial propagation release effect of Chinese sucker.     

Systemic immune responses to an inactivated,whole H9N2 avian influenza virus vaccine using class B CpG oligonucleotides in chickens

Commercial vaccines against avian influenza viruses (AIV) in chickens consist mainly of inactivated AIV, requiring parenteral administration and co-delivery of an adjuvant. Limitations in T helper 1 or T helper 2 biased responses generated by these vaccines emphasize the need for alternative, more efficacious adjuvants. The Toll-like receptor (TLR) 21 ligand, CpG oligodeoxynucleotides (ODN), has been established as immunomodulatory in chickens. Therefore, the objective of this study was to investigate the adjuvant potential of high (20 μg) and low (2 μg) doses of CpG ODN 2007 (CpG 2007) and CpG ODN 1826 (CpG 1826) when administered to chickens with a formalin-inactivated H9N2 AIV. Antibody responses in sera were evaluated in 90 specific pathogen free (SPF) chickens after intramuscular administration of vaccine formulations at 7 and 21 days post-hatch. Antibody responses were assessed based on haemagglutination inhibition (HI) and virus neutralization (VN) assays; virus-specific IgM and IgY antibody responses were evaluated by ELISA. The results suggest that the vaccine formulation containing low dose CpG 2007 was significantly more effective at generating neutralizing (both HI and VN) responses than formulations with high or low doses of CpG 1826 or high dose CpG 2007. Neutralizing responses elicited by low dose CpG 2007 significantly exceeded those generated by a squalene-based adjuvanted vaccine formulation during peak responses. A significantly higher IgM response was elicited by the formulation containing low dose CpG 2007 compared to high and low doses of 1826. Although the low dose of CpG 2007 elicited a higher IgY response than CpG 1826, the difference was not statistically significant. In conclusion, 2 μg of CpG 2007 is potentially promising as a vaccine adjuvant when delivered intramuscularly with inactivated H9N2 virus to chickens. Future studies may be directed at determining the mucosal antibody responses to the same vaccine formulations.     

Isolation and characterization of eleven novel microsatellite loci of Brachymystax lenok tsinlingensis, a threatened fish endemic to Shaanxi, China

Brachymystax lenok tsinlingensis is an endangered freshwater fish confined to cold-water mountain streams and rivers in Qinling Mountains of Shaanxi province, China. We isolated and characterized 11 microsatellite loci for Brachymystax lenok tsinlingensis and examined them for 42 individuals from Xushui River in Qinling Mountain. The number of alleles per locus ranged from 3 to 8, with an average of 5.3. The expected (He) and observed (Ho) heterozygosities were 0.3483–0.8874 and 0.4183–0.8421, respectively. Two loci deviated significantly from Hardy–Weinberg Equilibrium after Bonferroni correction. We expect these novel microsatellite markers from B. lenok tsinlingensis to be useful for population genetic conservation studies of the species.     

Isolation and characterization of twenty-six microsatellite loci for the tetraploid fish Dabry’s sturgeon (Acipenser dabryanus)

To evaluate the population genetic diversity as a means of devising conservation strategies, we constructed (CA)n and (CT)n enriched genomic libraries for Acipenser dabryanus, a narrowly endemic and endangered species in China. Twenty-six polymorphic microsatellite markers were developed using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Between 3 and 13 alleles per locus were observed. Mean expected heterozygosities (H _E) and Shannon-Wiener Diversity Indices (H′) per locus ranged from 0.303 to 0.806, and from 0.480 to 1.893, respectively. The microsatellite markers described here are valuable tools for the population genetics research of A. dabryanus.     

Development of microsatellite markers for Leptobotia rubrilaris, a threatened fish species in the Yangtze River

Leptobotia rubrilaris is a threatened fish species of the Yangtze River in China. A first set of sixteen polymorphism microsatellite loci isolated from this species was characterized. The number of alleles, observed and expected heterozygosity ranged from 2 to 8, 0.1667 to 1.000 and 0.1587 to 0.8342, respectively. Two loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction. The polymorphism information content ranged from 0.148 to 0.791. Ten loci were highly informative, four were moderately informative, and two was slightly informative. These polymorphic microsatellite markers will be useful to study genetic diversity and genetic structure of L. rubrilaris for conservation and management.     

白及幼根组织培养技术研究

目的：为白及组培快繁寻求最佳部位及最佳培养基配比。方法：研究不同激素浓度和配比对白及不同外植体诱导分化、增殖及生根的影响。结果：幼根最容易诱导出丛生芽,其最佳培养基为1/2MS＋1.0 mg·L-16-BA＋2.0 mg·L-1 NAA;丛生芽增殖最佳培养基为MS＋1.0 mg·L-16-BA＋0.05 mg·L-1 NAA;生根培养基以1/2MS＋0.5 mg·L-1 NAA为最佳。结论：建立了白及组培快繁体系。     

抗抑郁症胶囊中挥发油的提取工艺研究

目的：对中药方剂抗抑郁症胶囊中五味根茎类中药进行挥发油提取工艺的研究.方法：采用水蒸气蒸馏法提取挥发油,以挥发油的收率作为评价指标,以加水量、浸泡时间和提取时间为考察因素,用正交设计法来确定挥发油最佳提取工艺.结果：对五味中药进行提取试验,结果表明除石菖蒲以外其他四味中药材虽然含挥发油但挥发油含量太低,不适合工业化生产.最后只对含挥发油量最高的石菖蒲进行挥发油提取并开展单因素试验和正交试验确定最佳提取方工艺.结论：得到最佳提取工艺为选用石菖蒲饮片,浸泡2 h,加7倍水量,提取7 h,所得挥发油收率最高.     

Spatial spread and demographic expansion of Lyme borreliosis spirochaetes in Eurasia

The Lyme borreliosis (LB) group of spirochaetes currently comprises 18 named species that vary in their geographic distribution, host specificity and ability to cause disease in humans. In Europe three species are most abundant, Borrelia afzelii, Borrelia garinii and Borrelia valaisiana but only two of these (B. garinii and B. afzelii) are regularly found in Asia as well. A recently published study has shown that Borrelia species associated with birds, such as B. garinii, showed limited geographic structuring between European countries while, the rodent associated species, B. afzelii, showed extensive spatial structuring in Europe. Here, we use multilocus sequence analysis to show that when the wider, inter-continental, distribution is considered, there is evidence of spatial structuring even in the bird-associated species B. garinii. Furthermore, our investigations into historical LB populations provided evidence for range expansions of B. garinii and B. afzelii populations in Europe in the distant past. We propose that the expansion of B. afzelii in Europe may be linked to rodent population expansions after the last glacial maximum.