The protoxin Cry1Ac of Bacillus thuringiensis improves the protection conferred by intranasal immunization with Brucella abortus RB51 in a mouse model

Brucellosis is a zoonotic disease affecting many people and animals worldwide. Preventing this infection requires improving vaccination strategies. The protoxin Cry1Ac of Bacillus thuringiensis is an adjuvant that, in addition to increasing the immunogenicity of different antigens, has shown to be protective in different models of parasitic infections. The objective of the present study was to test whether the intranasal co-administration of pCry1Ac with the RB51 vaccine strain of Brucella abortus confers protection against an intranasal challenge with the virulent strain B. abortus 2308 in BALB/c mice. The results showed that co-administration of pCry1Ac and RB51, increased the immunoprotection conferred by the vaccine as evidenced by the following: (1) decrease of the splenic bacterial load when challenged intranasally with the virulent strain; (2) greater in vivo cytotoxic activity in response to the transference of previously infected cells; (3) further proliferation of cytotoxic TCD8+ cells in response to stimulation with heat-inactivated bacteria; (4) increased production of TNF-α and IFN-γ; and (5) significant IgG2a response. These results indicate that the use of the Cry1Ac protein as a mucosal adjuvant via the intranasal route can be a promising alternative for improving current RB51 vaccine against brucellosis.     

Protectivity conferred by immunization with intranasal recombinant outer membrane protein H from Pasteurella multocida serovar A:1 in chickens

Recombinant outer membrane protein H (rOmpH) is a potential fowl cholera vaccine candidate. The present study was aimed at developing rOmpH formulations for intranasal administration. The rOmpH was purified and formulated with either Escherichia coli enterotoxin B (LTB) or CpG oligodeoxynucleotides (ODN) as an adjuvant. Antibody responses in chickens intranasally immunized with rOmpH in combination with 2 different adjuvants were significantly increased (P<0.05) post immunization. Chicken survival rates showed that rOmpH formulated with ODN and LTB elicited 90% and 70% protection, respectively. Our findings indicated that rOmpH formulated with ODN elicited protection better than that formulated with LTB. Therefore, the vaccines formulations in the present study can be considered new intranasal vaccine formulations for fowl cholera in chickens.     

配方施肥对毛竹林新竹生长及经济效益的影响

通过"3414"施肥试验,研究了不同N、P、K配方施肥对福建三明毛竹林新竹生长及竹林经济产出的影响。结果表明,N、P、K不同配方对新竹的成竹率、新竹胸径与新竹产量以及对竹林经济产出的影响差异显著。14个配方处理中,N2P3K2的新竹数最高,为对照的1.69倍;N2P2K2的新竹胸径和产量均为最大,分别为对照的1.09和1.91倍,且新竹叶片的N、P、K、Mg浓度最为均衡。该试验条件下,施尿素540 kg/hm2、硫酸钾135 kg/hm2、过磷酸钙450~675 kg/hm2,能较为有效地促进新竹生长与产量持续。各配方施肥对毛竹林的增产幅度为18.1%~192.4%,增产效益明显,N2P2K2的竹林产值最高,是对照的2.92倍,资金产出率(产投比)前3位为N2P2K2、N2P3K2和N2P2K3,分别是3.83,3.43和3.26,N素对新竹生长的影响最大,缺N素、P素试验的资金产出率低于无施肥处理。     

黄波罗育苗中混雪层积时间及保留密度研究

对辽宁东部山区黄波罗播种育苗的混雪层积时间和保留密度进行了研究。结果表明:随混雪层积时间增加,黄波罗种子发芽率和出苗率呈上升趋势,混雪层积145d的发芽率和出苗率最大,分别达到81.35%、64.59%;1年生幼苗保留密度为100株·m-2时,Ⅰ级苗单位产量达到最高。     

移动通信基站的环境纠纷问题

指出了随着移动通讯的发展,移动通讯设施的不断增加,移动通信基站投诉纠纷问题逐渐凸显。根据浙江省辐射环境监测站2009～2013年期间承接的移动通信基站委托监测数据,分析了投诉纠纷的原因,并从公众、建设单位及环保部门的角度提出了应对投诉纠纷的意见及建议。     

Assessment of cross‐protection to heterologous strains of Flavobacterium psychrophilum following vaccination with a live‐attenuated coldwater disease immersion vaccine

Bacterial coldwater disease, caused by Flavobacterium psychrophilum, remains one of the most significant bacterial diseases of salmonids worldwide. A previously developed and reported live‐attenuated immersion vaccine (F. psychrophilum; B.17‐ILM) has been shown to confer significant protection to salmonids. To further characterize this vaccine, a series of experiments were carried out to determine the cross‐protective efficacy of this B.17‐ILM vaccine against 9 F. psychrophilum isolates (representing seven sequence types/three clonal complexes as determined by multilocus sequence typing) in comparison with a wild‐type virulent strain, CSF‐259‐93. To assess protection, 28‐day experimental challenges of rainbow trout (Oncorhynchus mykiss) fry were conducted following immersion vaccinations with the B.17‐ILM vaccine. F. psychrophilum strains used in challenge trials were isolated from several fish species across the globe; however, all were found to be virulent in rainbow trout. The B.17‐ILM vaccine provided significant protection against all strains, with relative percent survival values ranging from 51% to 72%. All vaccinated fish developed an adaptive immune response (as measured by F. psychrophilum‐specific antibodies) that increased out to the time of challenge (8 weeks postimmunization). Previous studies have confirmed that antibody plays an important role in protection against F. psychrophilum challenge; therefore, specific antibodies to the B.17‐ILM vaccine strain appear to contribute to the cross‐protection observed to heterologous strain. The ability of such antibodies to bind to similar antigenic regions for all strains was confirmed by western blot analyses. Results presented here support the practical application of this live‐attenuated vaccine, and suggest that it will be efficacious even in aquaculture operations affected by diverse strains of F. psychrophilum.     

牛源荚膜A型多杀性巴氏杆菌疫苗候选株的筛选及鉴定

为了筛选生长快、毒力强、免疫原性好、副反应小的牛源荚膜A型多杀性巴氏杆菌（Pasteurella multocida，Pm）灭活疫苗菌株，本试验选取6株来自不同地区致犊牛肺炎死亡的牛源荚膜A型多杀性巴氏杆菌分离株，测定了培养基生长曲线、小鼠毒力、菌体脂多糖（LPS）含量及各菌株灭活菌苗免疫小鼠和家兔后的抗体效价，并进行了攻毒保护试验。结果显示，分离株Pm2、Pm3、Pm5生长速度较快、毒力较强、LPS含量较多，均含有与毒力和免疫相关的ptfA和fimA基因；免疫小鼠及家兔未发现明显不良反应，在二免后14 d血清抗体达1：64~1：128，强毒攻毒后全部存活，而PBS对照组全部死亡。本试验结果表明，Pm2、Pm3、Pm5均可作为多杀性巴氏杆菌灭活菌苗的候选菌株，其中Pm3作为首选株。     

有机肥等氮替代化肥对红壤团聚体及玉米产量和品质的影响

为研究有机肥等氮替代化肥对土壤结构、玉米产量和品质的影响,设置有机肥等氮替代化肥的10%（M10）、20%（M20）、30%（M30）、40%（M40）,以单施化肥（M0）和不施肥（CK）为对照6个施肥处理,测定玉米不同生育期的土壤团聚体变化状况。结果表明：随有机肥等氮部分替代化肥比例的增加,玉米各生育期土壤>0.25mm水稳性团聚体（R_0.25）、平均质量直径（MWD）、几何平均直径（GWD）不断增加,团聚体破坏率（PAD）、分形维数（D）不断降低。与M0处理相比,M30和M40处理显著提高了玉米子粒产量,表现为M30>M40;M20、M30、M40处理较M0处理玉米子粒蛋白质含量显著增加了13.29%~25.03%,淀粉含量显著增加了39.84%~114.06%,可溶性糖含量显著增加了36.49%~54.05%;蛋白质含量表现为M30和M20处理显著大于M40处理,M30与M20处理间差异不显著;淀粉含量表现为M40>M30>M20,差异显著;可溶性糖含量表现为M40和M30处理显著大于M20处理,M30与M40处理间差异不显著。玉米子粒产量和品质与抽雄期、成熟期土壤团聚体R_0.25呈极显著正相关（P<0.01）,与PAD、D值呈极显著负相关（P<0.01）。因此,玉米生产中,利用有机肥等氮替代化肥培肥土壤,需兼顾玉米产量和品质,30%为适宜的有机肥等氮替代化肥比例。     

不同新型作用机制杀菌剂对梨树主要病菌毒力及防治流程应用

 为明确多种新型作用机制杀菌剂与引起梨树褐斑病、黑星病、白粉病等病原菌的有效对靶关系及制定梨树主要病害防治流程,采用菌丝生长速率法、离体叶片法与田间药效方法研究新型杀菌剂对靶标病原菌的毒力、对靶标病害的防效及其田间有效应用,建立替代梨树主要病害传统化学杀菌剂的防治流程技术。结果显示,双胍三辛烷基苯磺酸盐在离体叶片法下对褐斑病的防效大于85%,田间药效验证3次用药后7 d防效大于85%、30 d防效大于80%、90 d防效仍大于60%,兼治轮纹病菌其毒力EC₅₀值均小于1 μg·mL^-1。双胍三辛烷基苯磺酸盐、辛菌胺醋酸盐、吩嗪α-2羧酸在离体叶片法下对黑星病防效大于90%且毒力EC₅₀值小于1 μg·mL^-1;田间药效验证1次用药后7 d防效大于80%、30 d防效仍大于75%。噻肟菌酯、硝苯菌酯、丙硫菌唑在离体叶片法下对白粉病的防效大于85%;田间药效验证3次用药后7 d铲除效果大于70%、30 d仍大于60%。丙硫菌唑对黑斑、轮纹病菌毒力EC₅₀值均小于1 μg·mL^-1;吩嗪α-2羧酸同时对褐斑、黑斑、轮纹病菌毒力EC₅₀值小于1 μg·mL^-1。针对梨树主要病害发生期,选用新型杀菌剂替代传统杀菌剂制定防治技术流程,其综合防效达到88.94%。不同新型作用机制杀菌剂在防治梨树主要病害上的应用,能够降低抗药性产生,同时达到有效防治的目的。     

Protective immune responses of recombinant outer membrane proteins OmpF and OmpK of Aeromonas hydrophila in European eel (Anguilla anguilla)

Outer membrane proteins (Omps) of Gram‐negative bacteria have been proven to be efficient subunit vaccines against bacteriosis. In this study, OmpF and OmpK of Aeromonas hydrophila were expressed, and their immune protective effects in European eel (Anguilla anguilla) were evaluated. The genomic DNA of A. hydrophila 322A was used as a template, and two kinds of prokaryotic expression plasmids, pET‐32a‐OmpF and pET‐32a‐OmpK, were constructed. Recombinant OmpF protein (r‐OmpF) and r‐OmpK were purified and were proven to have antigenicity by Western‐blot analysis. r‐OmpF and r‐OmpK were used as immunogens to immunize European eel by intraperitoneal injection. The mRNA expression of 6 immune‐related genes (IgM, IL‐10, IRF3, IRF7, LysG4 and HexB) in the liver tissues of eels at 1 hr, 3 hr, 6 hr, 12 hr, 24 hr, 72 hr and 10 days postimmunization was analysed by real‐time PCR. At 30 dpi, the serum antibody response was measured by ELISA. Fish were attacked at 15 dpi by live 322A to assess the protective immunity of r‐OmpF and r‐OmpK. All the six tested genes responded to r‐OmpF or r‐OmpK vaccination at varying degrees. The serum antibody titre of r‐OmpF‐ and r‐OmpK‐immunized groups was 1:1,600 and 1:3,200 respectively. In addition, r‐OmpF gave 35.5% of the relative immune protection rate to European eels, while r‐OmpK gave 70.0%. By analysing the protective immunity and the regulatory role in the immune‐related gene expression of the two recombinant proteins that were studied, it was found that r‐OmpK was a potential vaccine candidate against A. hydrophila.