Characterization of Sarcocystis species in domestic animals using a PCR-RFLP analysis of variation in the 18S rRNA gene: a cost-effective and simple technique for routine species identification

Thirteen restriction endonucleases were used to investigate nucleotide sequence variation in the 18S rRNA DNA of 88 individuals from ten Sarcocystis taxa collected as cysts from their intermediate hosts, swine, cattle and water buffalo. A DNA sequence of

Identification and characterization of a novel envelope protein in Rana grylio virus

Viral envelope proteins have been proposed to play significant roles in virus infection and assembly. In this study, an envelope protein gene, 53R, was cloned and characterized from Rana grylio virus (RGV), a member of the family Iridoviridae. Database searches found its homologues in all sequenced iricloviruses, and sequence alignment revealed several conserved structural features shared by virus capsid or envelope proteins: a myristoylation site, two predicted transmembrane domains and two invariant cysteine residues. Subsequently, RT-PCR and Western blot detection revealed that the transcripts encoding RGV 53R and the protein itself appeared late during infection of fathead minnow cells and that their appearance was blocked by viral DNA replication inhibitor, indicating that RGV 53R is a late expression gene. Moreover, immunofluorescence localization found an association of 53R with virus factories in RGV-infected cells, and this association was further confirmed by expressing a 53R-GFP fusion protein in pEGFP-N3/53R-transfected cells. Furthermore, detergent extraction and Western blot detection confirmed that RGV 53R was associated with virion membrane. Therefore, the current data suggest that RGV 53R is a novel viral envelope protein and that it may play an important role in virus assembly. This is thought to be the first report on a viral envelope protein that is conserved in all sequenced iridoviruses.

Genomic organization, gene duplication, and expression analysis of interleukin-1 beta in channel catfish (Ictalurus punctatus)

Interleukin-1 beta (IL-1 beta) is one of the pivotal early response pro-inflammatory cytokines that enables organisms to respond to infection and induces a cascade of reactions leading to inflammation. In spite of its importance and two decades of studies in the mammalian species, genes encoding IL-1 beta were not identified from non-mammalian species until recently. Recent research, particularly with genomic approaches, has led to sequencing of IL-1 beta from many species. Clinical studies also Suggested IL-1 beta as an immunoreagulatory molecule potentially useful for enhancing vaccination. However, no IL-1 beta genes have been identified from channel catfish, the primary aquaculture species from the United States. In this study, we identified two distinct cDNAs encoding catfish IL-1 beta. Their encoding genes were identified, sequenced, and characterized. The catfish IL-1 beta genes were assigned to bacterial artificial chromosome (BAC) clones. Genomic studies indicated that the IL-1 beta genes were tandemly duplicated on the same chromosome. Phylogenetic analysis of various IL-1 beta genes indicated the possibility of recent species-specific gene duplications in channel catfish, and perhaps also in swine and carp. Expression analysis indicated that both IL-1 beta genes were expressed, but exhibited distinct expression profiles in various catfish tissues, and after bacterial infection with Edwardsiella ictaluri. (c) 2005 Elsevier Ltd. All rights reserved.

施肥措施对黄土旱塬区小麦产量和土壤有机碳积累的影响

利用中国科学院长武农田生态试验站的长期田间试验(1984年2~007年),研究了小麦产量,耕层有机碳变化,评价了土壤管理和气候因素对土壤有机碳(Soil organic C,SOC)变化的影响。研究涉及6个处理:休闲地(F);不施肥(CK);有机肥(M);氮肥(N);氮、磷肥(NP)和氮、磷、有机肥(NPM)处理。结果表明,施肥可以显著提高作物产量和SOC积累,CK、M、N、NP、NPM处理平均产量依次为1.5、2.6、2.0、3.3、4.0 t/hm2,2007年F、CK、M、N、NP、NPM处理0—20 cm土层SOC积累量依次为-1.09、0.76、8.59、1.02、3.42和9.5 t/hm2。作物产量与SOC含量呈显著的正相关关系(r=0.80),有机碳输入量与SOC含量相关性更好(r=0.97),外源有机碳的输入也是提高SOC的重要措施。施肥措施对作物固碳和SOC影响存在显著(P<0.05)差异。土壤固碳速率(Y)与SOC输入量(X)符合线性方程Y=0.231X-0.0813(r=0.98)。施肥可以提高黄土高原半干旱地区土壤生产力和SOC的积累,且无机肥和有机肥配施效果最佳。

施肥对黄土旱塬区黑垆土土壤肥力及硝态氮累积的影响

本文以长期定位试验为依托,研究了黄土高原旱塬区黑垆土大田对比试验和长期定位施肥对土壤肥力及硝态氮累积和淋溶的影响。结果表明:长期施用有机肥能够明显增加土壤养分,氮磷和有机肥配施效果显著;和1984年土壤养分状况相比,大田对比试验土壤有机质增加了27.1%,全氮和全磷提高了84.2%和34.8%,有效氮、有效磷和速效钾增加了46.9%、540.0%和10.2%,养分水平与长期定位试验中氮磷配施相近。长期定位试验中氮磷配施或与有机肥配施能够有效地减少土壤剖面中硝酸盐的累积和淋溶,氮肥单施硝态氮累积量最大,为1006.4kg/hm2,大田对比试验土壤硝态氮总累积量较长期定位试验中施用氮肥处理的总累积量少。

黄土丘陵区免耕条件下两种土壤酶活性变化

为了确定合理环保的耕作制度,2007~2008安塞田间定位试验黄土丘陵旱作农区大豆(Glycine maxL)、玉米(Zea maysL)、红小豆(Semen Phaseoli)、马铃薯(Solanum tuberosumLinn.)在翻耕化肥(CF)、翻耕有机肥(CM)、翻耕无肥(CN)、免耕化肥(NF)、免耕有机肥(NM)、免耕无肥(NN)等水平下的农田土壤脲酶、蔗糖酶活性。结果表明:在作物花期,大豆、玉米土壤脲酶活性较高,蔗糖酶活性较低,而红小豆、马铃薯则与之相反,差异极显著。到作物收获后,玉米土壤脲酶、蔗糖酶活性增高,增幅在83%以上,而马铃薯、红小豆、大豆三种作物土壤脲酶活性降低,降幅在10%以上,蔗糖酶活性增强,增幅在40%以上。从花期到收获后,免耕降低了土壤脲酶活性,提高了土壤蔗糖酶活性,在黄土丘陵沟壑旱作农区两种土壤酶活性表现较优的作物为大豆、玉米,较优的处理为NM玉米、NF大豆,其田间环保效应为:作物生长期间两种酶活性比较高,收获后两种酶活性则降低,有利于提高作物生长期土壤肥效利用率,减少作物收获后温室气体的排放。

黄土高原旱地小麦氮磷钾与有机肥优化配施试验

试验研究了不同施肥处理对小麦产量及肥料、水分利用率的影响。结果表明:氮磷配施对产量的贡献率最高,产量达5 099.8 kg/hm2,增产率达21.5%,较氮、磷肥单施产量分别增产14.0%和18.4%,具有正交互效应;单施有机肥较NM、PM、NPM增产率分别达到6.63%、4.12%、11.3%。化肥配施可显著提高旱地小麦的肥料利用率和水分利用效率,氮磷有机肥配施肥料氮、磷肥的利用率分别较氮、磷肥单施提高1.71和12.55个百分点;氮磷配施可同时提高氮肥、磷肥肥效,此时氮贡献率为15.53%,磷贡献率达12.26%,且水分利用率提高了24.1%,耗水系数降低了17.57%。

有机肥对设施菜地土壤-植物系统氮运移的影响

在设施菜地条件下，研究了不同有机肥施入量在黄瓜生长期对土壤-植物系统NO3--N迁移累积的影响。结果表明，黄瓜生长期土壤-植物系统NO3--N的迁移累积规律受黄瓜生长期、有机肥施用水平和土壤肥力的影响。盛果期，不同肥力土壤各个土层的硝酸盐含量和黄瓜叶片及体内的硝酸盐含量均高于黄瓜的其它生长期；对于不同的施肥水平，当施肥量为60 t hm-2时各个土层的不同肥力土壤硝酸盐含量均高于其它处理，且高肥力土壤条件下黄瓜体内的硝酸盐含量在黄瓜生长旺盛期超过国家安全食品标准(410mgkg-1)，当施肥量低于20 t hm-2时不同肥力土壤各个土层未出现硝酸盐显著累积现象，且在黄瓜生长的各个时期，黄瓜体内的硝酸盐含量均未超标。 土壤硝酸盐的垂直运移过程受有机肥施用水平和土壤肥力的影响，施肥量低于20 t hm-2时不同肥力土壤各土层未出现硝酸盐显著累积现象，当施肥量为60 t hm-2时土壤硝酸盐累积峰值随土壤肥力水平的提高而增加；40-150 cm土壤剖面中硝酸盐累积总量随有机肥施用量的增加而增加；不同肥力土壤在相同有机肥施用水平下40-150 cm土壤硝酸盐累积总量的变化量不同，土壤肥力越高土壤硝酸盐累积总量的绝对增加量越大，说明相同的有机肥处理对高肥力土壤硝酸盐累积影响更大。

有机肥对设施菜地土壤磷素累积及有效性影响

本文以沈阳城郊设施菜地为研究对象，系统研究了不同有机肥处理(0-60 t hm-2)对不同本底肥力设施菜地土壤磷素累积及有效性的影响。结果如下： 施有机肥各处理土壤0-20cm全磷含量与CK之间均达显著差异，且施有机肥可显著增加0-20cm土层的活性有机磷、中等活性有机磷和中稳性有机磷的累积量。其中以中等活性有机磷和中稳性有机磷含量增加较显著。不同本底磷肥力土壤活性有机磷、中等活性有机磷和中稳性有机磷均与土壤速效磷呈极显著正相关关系。 施用有机肥各处理土壤0-20cm速效磷的含量随有机肥施用量的增加而明显增加。土壤速效磷含量在黄瓜不同生育期的变化规律是 ：黄瓜开花期达最高，坐果期和拉秧期有所下降。 对于土壤本底磷肥力达1.97g kg-1的设施菜地，有机肥施用量10t hm-2可基本保持施肥当季土壤0-20cm速效磷水平；施用有机肥20t hm-2可保持土壤本底磷肥力0.86g kg-1的设施菜地0-20cm土壤速效磷水平，如过量施用有机肥则可进一步提高土壤全磷和速效磷含量，进而增加磷淋失的环境风险。 对于不同本底磷肥力设施菜地施用相应适量有机肥可提高黄瓜产量，并保持土壤磷肥力水平。过量施用有机肥则不仅增产效应不明显，且随有机肥施用量的增加，磷素表观利用率降低，磷淋失环境风险增加，经济效益降低。

长期施肥对农田土壤N、P转化酶活性的影响

本试验利用国家黑土肥力与肥料效益长期定位监测基地为平台，研究了多种施肥方式（包括休闲、不施肥、氮、氮钾、氮磷钾、秸秆配施化肥、猪粪配施化肥等）对参与土壤氮转化过程的蛋白酶 、脲酶、硝酸还原酶、氨氧化酶以及参与土壤有机磷矿化的磷酸三酯酶、磷酸二酯酶和磷酸单酯酶活性和动力学特性的作用，以探讨不同施肥方式对氮磷转化酶活性的影响，得出如下结果： 1. 不同施肥处理对土壤氮转化酶促过程影响不同，有机肥配施化肥处理显著增强了蛋白酶、脲酶、氨氧化酶活性。有机肥配施化肥处理土壤速效氮含量高于化肥处理，硝态氮积累量较大，铵态氮含量差异不显著，有机肥配施化肥能够显著促进尿素水解和硝态氮的积累，表明有机肥配施化肥有机氮的矿化强度及硝化作用强于化肥处理。施肥对硝酸还原酶活性没有显著影响。相关分析表明，土壤蛋白酶、脲酶和氨氧化酶活性与微生物量碳呈显著正相关，氨氧化酶活性与脱氢酶呈显著正相关。 2. 有机肥配施化肥处理显著提高了土壤速效磷的含量，增强了土壤磷酸二酯酶和中性磷酸单酯酶活性，土壤速效磷含量过高的情况下，速效磷对中性磷酸单酯酶产生一定程度的抑制作用。与对照相比，化肥处理显著增高了中性磷酸单酯酶活性，但是对磷酸二酯酶有一定的抑制作用。相关分析表明，土壤磷酸二酯酶活性与土壤脱氢酶、速效磷呈显著正相关，中性磷酸单酯酶与速效磷含量呈负相关。

有机物料改良滨海盐土的肥盐效应研究

本文通过微区、盆裁和根际试验相结合的方法，研究了有机物料对黄河三角洲东营滨海盐土的盐分、养分动态和酶活性的影响，结果表明，盆裁和微区试验土壤盐分随稻草粉用量增大而递增，主要在于二价离子的上升；土壤交换性钙、镁、钾上升，钠下降；土壤pH略有降低，土壤速效氮、磷、钾养分明显上升。有机物料对土壤铁、锌活化作用明确，而对锰、铜影响不大。有机物料能增强土壤转化酶、碱性磷酸酶和脲酶的活性，从而对提高土壤有机质矿化的速率，以及改善作物的根际养分状况具有良好作用。

耐酸产甲烷菌的分离及其复合菌剂的研究

本文从成都龙泉垃圾填埋场和宜宾造纸厂分离到耐酸性能优良的高温产甲烷菌RY3和中温产甲烷菌SH4，并将其与实验室现有的利用不同底物的产甲烷菌配伍组合成了复合菌剂。采用活性污泥作为固体附着物，研制出了固体产甲烷菌复合菌剂。 菌株RY3的pH耐受范围为5.5～10.5，最适生长pH 6.0～8.0。菌株RY3为革兰氏阳性，长杆状，多数单生，不运动；菌落浅黄色，形状近圆形；利用H2+CO2或甲酸盐作为唯一碳源生长，不利用乙酸盐，对氯霉素非常敏感。该菌最适生长温度为55℃～65℃，最适NaCl浓度为0～2%。根据形态和生理生化特性及16S rDNA序列分析将其初步定为热自养甲烷热杆菌（Methanothermobacter thermautotrophicus）。添加RY3菌液与仅添加厌氧污泥作为接种物相比一周内可使达到最大产甲烷速率所需时间缩短三分之二，甲烷总产量提高约1.8倍。菌株SH4的生长pH范围5.5～9.5，其对酸碱具有良好的适应性，培养3天后，在初始pH值为6.0～8.0的培养基中甲烷产量相差不大，且基本达到最大产量。SH4革兰氏染色阳性，短杆状，多数单生，不运动；菌落近圆形，微黄；利用H2+CO2或甲酸盐作为唯一碳源生长，不利用乙酸盐，对氯霉素非常敏感。SH4最适生长pH 为7.0，最适生长温度为35℃，最适NaCl浓度为0～1.5%。实验表明，添加SH4菌液与仅添加厌氧污泥作为接种物相比可使产甲烷启动时间缩短三分之一，甲烷总产量亦有大幅提高。从形态和生理生化特征以及16S rDNA序列分析表明SH4为嗜树木甲烷短杆菌（Methanobrevibacter arboriphilus）。 以活性污泥为附着物，与培养基和菌种经搅拌后厌氧发酵可得产甲烷菌固体复合菌剂。固体复合菌剂的pH耐受范围为5.5～9.5，温度耐受范围为15℃～65℃，表明其对环境的适应性较强。以猪粪为底物进行厌氧发酵，接种复合菌剂进行试验，以接种实验室长期富集的产甲烷厌氧污泥作为对照，在20℃时，发酵甲烷浓度与对照基本一致，但每日产气量优于对照，第15天时接种复合菌剂的发酵瓶每日产气量是对照的1.59倍；50℃时达到最大甲烷含量所需时间比对照缩短三分之二，三周内总产气量约为对照的2.7倍，甲烷总产量约为2.8倍。以不加接种物为对照，接种复合菌剂20℃时发酵甲烷含量达到50%约需2周，对照2周内甲烷含量最高仅为4.3%；50℃时接种复合菌剂发酵仅需约1周甲烷含量便可达50%，对照则至少需要2周。 In this paper, high-temperature Methanogen RY3 and middle-temperature SH4 were isolated from Chengdu Longquan refuse landfill and Yibin paper mill. They could be used to make compound inoculum that producing methane with the existing Methanogens utilized different substrate. With using anaerobic activated sludge be solid fixture, the process had been designed to produce solid compound inoculum. Strain RY3 possessed excellent capacity of acid and alkali-tolerant. The pH-tolerant scale of RY3 was 5.5～10.5 and its optimum pH value for growth was 6.0～8.0. RY3 was G+, long-rod shape, monothetic and nonmotile, the colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by RY3 as sole C-source, and it was very sensitive to chloramphenicol. Besides, strain RY3 grew fastest at 55℃～65 and 0℃～2% NaCl. Characteristics of modality and physiology with sequence analysis of the 16s rDNA gene of strain RY3 preliminarily showed that it was Methanothermobacter thermautotrophicus. The experiments indicated that the time which began to produce methane with the highest velocity could be shortened two third by adding RY3 in one week, and the total methane production also was 1.8 times than before. Strain SH4 possessed wide scale of growing pH（5.5～9.5）and excellent ability of acclimatizing itself to acid-alkali. The methane production had no apparent difference among those cultivated in different initial pH（6.0～8.0）after three days and equaled to the maximum production basically. Cells of SH4 were G+, short-rod sharp, monothetic and nonmotile. The colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by SH4 as sole C-source, and it was very sensitive to chloramphenicol. Besides, it grew fastest at pH 7.0，55 ℃～65 and 0℃～2% NaCl concentration. The experiment indicated the time that began to produce methane could be shortening one third by adding SH4. And the total methane production also rose apparently. Characteristic of modality and physiology with sequence analysis of the 16S rDNA gene of strain SH4 demonstrated it was Methanobrevibacter arboriphilus. The activated sludge was utilized as fixture, mixed with culture medium and inocolum, that the solid compound inoculum could be produced by anaerobic fermentation. The compound inoculum could grow between pH 5.5～9.5, 15℃～65. It demonstrated the compound inoculum ha℃ve great ability of adapting to circumstance. In the experiment that making pig manure be substrate and taking the anaerobic sludge producing methane that cultured in long term in laboratory to be comparison, the concentration of methane in fermentation added compound inoculum almost equal to the comparison at 20℃, but the volume of gas production could be a little higher. The gas production everyday inoculated compound inoculum was 1.59 times to comparison. The time that the concentration of methane to maximum could be shortening by two third by adding compound inoculum, and the total gas production was 2.7 times to comprison while the total methane production was 2.8 times. If take the no inoculum be the comprasion, anaerobic fermentation added compound inoculum made the concentration of methane to 50% in 2 weeks but the comparison only to 4.3% at 20℃. The time that the concentration of methane to 50% by adding compound inoculum only need 1 week, but the comparison need 2 weeks at 50℃.

Influence of Hypericum perforatum Extract on Piglet Infected with Porcine Respiratory and Reproductive Syndrome Virus

To study the influence of Hypericum perforatum extract (HPE) on piglets infected with porcine respiratory and reproductive syndrome virus (PRRSV), enzyme-labeled immunosorbent assay (ELISA) and cytopathic effect (CPE) were used to determine in vitro whether HPE could induce swine pulmonary alveolar macrophages (PAMs) to secrete IFN-gamma, and whether PRRSV titers in PAMs were affected by the levels of HPE-induced IFN-gamma. HPE (200 mg kg(-1)) was administrated by oral gavage to piglets infected with the PRRSV in vivo to observe whether HPE affected the viremia, lung viral titers, and weight gain of piglets infected with PRRSV. The results showed that HPE was capable of inducing PAMs to produce IFN-gamma in a dose dependent manner and HPE pretreatment was capable of significantly reducing PRRSV viral titers in PAMs (P<0.01). Administration of HPE to the PRRSV-infected animals significantly (P<0.05) reduced viremia over time as compared with the PRRSV-infected animals. But there was not significant decrease in lung viral titers at day 21 post-infection between the HPE-treated animals and the PRRSV-infected control piglets. There were no significant differences in weight gain over time among the HPE-treatment animals, the normal control, and the HPE control animals. The PRRSV-infected animals caused significant (P<0.01) growth retardation as compared with the HPE controls and the normal piglets. It suggested that HPE might be an effective novel therapeutic approach to diminish the PRRSV-induced disease in swine.

氮和磷在饲养-堆腐环中的循环率及有机肥料养分利用率

A five-year experiment indicated that the average loss rate of N and P in harvested products through a feeding-composting cycle was about 50% and 15%,respectively.Under high yield condition,the amount of recycled N and P from 80% harvested products and through a feeding-composting cycle in farming systems was about 3751 and 814 kghm~(-2),equivalent to 25%34% of N and 32%56% of P from chemical fertilizers applied each year to the systems.The apparent recoveries of N and P in organic manure increased with the prolongation of fertilization,indicating a synergetic residual effect existed,and was 61% and 39% in average in the five-year experiment.The use of nutrients recycled in the farming systems could not only improve soil fertility,but also increase the recoveries of nutrients and reduce the use of chemical fertilizers.

不同水肥条件对马铃薯肥料N利用率的影响

在旱棚控制条件下 ,用五因素五水平二次回归正交旋转组合设计 ,研究了水肥施用分配 ,补水量 ,施N量 ,施K量 ,有机肥施用量的综合作用效应 .结果表明 ,五因素对肥料N利用率的作用顺序为施N量 >补水量>水肥施用分配 >有机肥施用量 >施K量 .水肥施用分配相对于不同水、化肥 (N、K )、有机肥量的选择 ,可以大大地提高肥料N利用率 ;量少应重前施用 ,量多重后施或均施为好 .补水量与施N量的耦合规律为 :高水高肥效果最佳 ,肥料N利用率可达 5 7 83 % ,中水中N及低水低N次之 ,为 2 9 17%～ 40 99% ;高水低N或低水高N导致最低的利用率 ,为 2 2 87%或 2 2 5 1%