Physicochemical and Biological Characterization of Fucoidan from Fucus vesiculosus Purified by Dye Affinity Chromatography

A comparative study concerning the physicochemical, monomeric composition and biological characters among different fucoidan fractions is presented. Common purification techniques for fucoidan usually involve many steps. During these steps, the important structural features might be affected and consequently alter its biological activities. Three purified fractions were derived from Fucus vesiculosus water extract which, afterwards, were purified by a recently-developed dye affinity chromatography protocol. This protocol is based on dye-sulfated polysaccharide interactions. The first two fractions were obtained from crude precipitated fucoidan at different pH values of the adsorption phase: pH 1 and 6. This procedure resulted in fucoidan_1 and 6 fractions. The other, third, fraction: fucoidan_M, however, was obtained from a buffered crude extract at pH 1, eliminating the ethanol precipitation step. All of the three fractions were then further evaluated. Results revealed that fucoidan_M showed the highest sulfur content (S%), 12.11%, with the lowest average molecular weight, 48 kDa. Fucose, galactose, and uronic acid/glucose dimers were detected in all fractions, although, xylose was only detected in fucoidan_1 and 6. In a concentration of 10 µg·mL⁻¹, Fucoidan_6 showed the highest heparin-like anticoagulant activity and could prolong the APTT and TT significantly to 66.03 ± 2.93 and 75.36 ± 1.37 s, respectively. In addition, fucoidan_M demonstrated the highest potency against HSV-1 with an IC₅₀ of 2.41 µg·mL⁻¹. The technique proved to be a candidate for fucoidan purifaction from its crude extract removing the precipitation step from common purification protocols and produced different fucoidan qualities resulted from the different incubation conditions with the immobilized thiazine toluidine blue O dye.     

Frequency of occupation with different simultaneously offered devices by fattening pigs kept in pens with or without straw

The aim of this study was to investigate to what extent different occupational devices (pendular beam, cross of chains or lifting beam) offered simultaneously were used by one or more pigs kept in pens with or without straw during the fattening period. The data were obtained in four fattening periods. In every round four pens with nine animals each were available, two of them had a solid concrete floor and were littered with straw the other two had a partially slatted floor without any straw. At three stages during each fattening period the behaviour of the pigs was video-recorded for 24 h and their use of each occupational tool was analysed second-wise.In both housing systems the cross of chains was used longer for occupation than the other devices. The pendular beam was used least of all. All three devices could have been used by more than one pig at the same time, but the cross of chains was used longer by two to four pigs simultaneously than the others. It was observed that pigs in pens without straw used the cross of chains and the lifting beam significantly (p < 0.05) longer than the pigs in the pens with straw.All three devices are suitable to enrich a housing system without any litter. Especially a cross of chains allows the pigs to perform their explorational behaviour.     

Development of two microsatellite multiplex PCR systems for high throughput genotyping in Populus euphratica

Eighteen microsatellite primer pairs previously developed at Oak Ridge National Laboratory for Populus tremuloides Michx. and Populus trichocarpa Torr. & Gray were screened for amplification in Euphrates poplar, Populus euphratica Oliv. Thirteen loci were found to express polymorphisms ranging from two to 17 alleles. The eight most variable loci were selected to set up and optimize two multiplex polymerase chain reaction (PCR) assays. Three populations containing altogether 436 trees were used to characterize the selected loci and ascertain their applicability for parentage analysis and genotyping studies. Through cross-checking of clonal identity against sex of the genotyped trees we estimated the maximum error rate for merging genotypes to be less than 0.045. Foundation project: This study was supported by the Deutsche Forschungsgemeinschaft DFG (grant number SCHN 1080/1-1)     

两种微卫星多样PCR系统应用于胡杨高效生产基因型

对已在橡树岭国家实验室建立起来的美洲山杨(Populus tremuloides Michx.)和毛果杨(Populus trichocarpa Torr.＆ Gray)18个微卫星引物进行筛选,用于胡杨相关基因的扩增.发现有13个基因位点可以表达基因多态性,范围是2～17个等位基因.选择了8个最易变的基因位点,来构建和优化2种多样PCR法.用来自3个种群的436株树木表征选择的位点,确定他们在亲缘关系分析、基因型研究的适用性.通过克隆基因型鉴定树木性别的交叉检验,我们估计并入基因型最大误差率小于O.045.     

Characterization of protection against systemic infection and disease from experimental bovine viral diarrhea virus type 2 infection by use of a modified-live noncytopathic type 1 vaccine in calves

OBJECTIVE: To evaluate protection resulting from use of a modified-live noncytopathic bovine viral diarrhea virus (BVDV) type 1 vaccine against systemic infection and clinical disease in calves challenged with type 2 BVDV. ANIMALS: 10 calves, 5 to 7 months of age. PROCEDURES: Calves were allocated (n = 5/group) to be nonvaccinated or vaccinated SC on day 0 with BVDV 1 (WRL strain). Calves in both groups were challenged intranasally with BVDV type 2 isolate 890 on day 21. Rectal temperatures and clinical signs of disease were recorded daily, and total and differential WBC and platelet counts were performed. Histologic examinations and immunohistochemical analyses to detect lesions and distribution of viral antigens, respectively, were performed. RESULTS: After challenge exposure to BVDV type 2, nonvaccinated calves developed high rectal temperatures, increased respiratory rates, viremia, leukopenia, lymphopenia, and infection of the thymus. Vaccinated calves did not develop high rectal temperatures or clinical signs of respiratory tract disease. Vaccinated calves appeared to be protected against systemic replication of virus in that they did not develop leukopenia, lymphopenia, viremia, or infection of target organs, and infectious virus was not detected in peripheral blood mononuclear cells or the thymus. CONCLUSIONS AND CLINICAL RELEVANCE: The modified-live BVDV type 1 vaccine protected against systemic infection and disease after experimental challenge exposure with BVDV type 2. The vaccine protected calves against infection and viremia and prevented infection of target lymphoid cells.     

Impact of rank position on fertility of sows

The investigations were carried out with 484 sows from two farms (farm A: housing the sows in small groups of 8 animals each, farm B with a large group of 100 sows) and a total number of 982 inseminations. The number of agonistic interactions was registered for each sow during 48 h after mixing soon after weaning the piglets at farm A. The individual rank place in the social hierarchy was calculated on the basis of wins and defeats and the sows were divided in high and low ranking sows. At farm B the rank position was estimated on the basis of the daily feeding order at two electronic feeding stations (first half of the sows in the feeding order = high ranking, second half = low ranking). Additionally, the following parameters were recorded for each sow: parity, genotype, farrowing rate and litter size (total and alive born piglets). The analysis showed that sows with a high rank position had a significantly higher farrowing rate (88.8%) compared to group-mates with low rank places (82.8%, p = 0.051) (farm A). Sows with a high rank position reached a significantly higher litter size of total born piglets (12.66, 16.14 piglets per litter respectively) than the low-ranking group-mates (12.13, 14.83 piglets/litter respectively — farms A and B). When mixing sows, the time and the conditions (e.g. group size, space allowance per sow) have to be considered to prevent the negative influence of low rank order on fertility.     

中国家鸡和红色原鸡mtDNA控制区遗传多态性及系统进化分析

通过线粒体DNA控制区的结构和多态性来研究中国家鸡和红色原鸡的遗传多态性与系统进化。测定14个中国地方鸡种和红色原鸡2个亚种的256个个体线粒体DNA控制区部分序列约560bp,结果表明,A、C、G、T这4种核苷酸的平均比例分别为25．∞％、37．40％、4．40％和33．20％。共发现44个变异位点,约占分析位点总数的7．86％,没有观测到插入／缺失,颠换和转换之比为0．13;共具有32种单倍型,9种为共享单倍型;16个群体内单倍型多样度从0到0．964,单倍型变异度总体为0．909&#177;0．014,整体的平均核苷酸差异数为7．276,核苷酸多样度为1．851％。群体间核苷酸分歧度（Dxy）在0．747％～3．125％之间变化,核苷酸净遗传距离（Da）为0．015％～2．633％。16个群体表现出较高水平的遗传多态性,群体间表现出显著的遗传分化。群体遗传多态性和亲缘关系分析表明,一些中国家鸡的群体（如固始鸡和仙居鸡）起源于泰国红色原鸡Gallus gallu sgallus亚种,一些中国家鸡的群体（如茶花鸡和藏鸡等）起源于中国红色原鸡Gallus gallus spadiceus亚种,在一些中国地方鸡种还同时具有这2种红色原鸡的遗传贡献;认为中国家鸡起源于泰国或单纯起源于中国的观点都是不全面的。     

Field efficacy of ivermectin plus praziquantel oral paste against naturally acquired gastrointestinal nematodes and cestodes of horses in North America and Europe

The efficacy of an oral formulation of ivermectin plus praziquantel in the reduction of nematode and cestode egg counts in horses was assessed in 273 horses under field conditions at 15 sites in North America (n = 6) and Europe (n = 9). Horses were confirmed by fecal examination to have natural infections of strongyles (100%) and tapeworms (76%). Replicates of four horses were formed at each site, and in each replicate three animals received ivermectin (0.2 mg/kg body weight) plus praziquantel (1 mg/kg body weight) oral paste and one animal remained untreated or received vehicle paste. Fecal samples were collected for fecal nematode and cestode egg counting before and 7, 8, 9, 14, 15, and 16 days after treatment. Horses treated with ivermectin plus praziquantel oral paste had significantly (P <.01) lower posttreatment strongylid and cestode egg counts (reductions of 98% or more) than controls. Combined site analyses revealed that 95% or 96% of the horses positive for cestode eggs before treatment that were treated with ivermectin plus praziquantel were negative for cestode eggs at each posttreatment fecal examination. No adverse reactions attributable to ivermectin plus praziquantel oral paste treatments were observed. The results of the studies demonstrated that ivermectin plus praziquantel paste was highly effective in reducing egg shedding by gastrointestinal nematodes and cestodes, and no adverse reactions were observed in horses treated under field conditions.