一株野鸟源H9N2亚型禽流感病毒的全基因组序列分析及致病性研究

为进一步了解2014年分离自我国南方野鸟粪便中的一株H9N2亚型禽流感病毒(AIV)Wide Bird/Hu N/SC1400/2014(H9N2)(WB/400/14)的生物学特性,本研究对其进行全基因组序列测定、进化分析及SPF鸡、SPF鸭和BALB/c小鼠的感染性试验。序列分析显示:该分离株的HA裂解位点基序为333PAASDR↓GL340,其中不存在多个连续的碱性氨基酸,符合低致病性禽流感病毒(LPAIV)氨基酸序列特征。该分离株不同基因片段来源较复杂,分别与H9、H6、H4、H1、H11、H10、H3等多种亚型的LPAIV同源性较高,呈现明显的多样性。感染性试验显示,WB/400/14不能够在SPF鸡和小鼠体内有效复制,但病毒感染SPF鸭后能够在部分脏器中检测到病毒的存在,并且感染鸭能通够过咽喉和泄殖腔同时向外排毒,而同居感染鸭仅通过泄殖腔向外排毒,表明分离株在SPF鸭群中具有良好的水平传播能力。本研究为AIV的监测和防控提供实验依据。     

2株鸭源 H11N9亚型禽流感病毒的基因遗传进化及致病性分析

为了解从湖南省洞庭湖区鸭群中分离的2株 H11N9亚型禽流感病毒变异特点、进化规律及生物学特性,本研究对2株H11N9亚型禽流感病毒的HA、NA序列进行同源性和遗传进化分析,并用2株毒株对SPF鸡进行致病性试验。结果显示,本试验分离到2株 H11N9亚型禽流感毒株的 HA裂解位点均没有多个连续的碱性氨基酸插入,属于低致病性毒株;HA基因的受体结合位点均非常保守,具有典型的禽源性特征;NA基因序列与在周边国家野鸟中分离的H11N9亚型毒株的氨基酸同源性较高;鼻腔接种SPF鸡后,均能使鸡感染并通过喉头或泄殖腔排毒,但感染的鸡均不表现明显的临床症状,并且不能使同居鸡感染排毒。     

Cloning and Sequence Analysis of N and M Genes of Avian Infectious Bronchitis Virus Guangxi Strain

To investigate genetic variation of avian infectious bronchitis virus (IBV) in Guangxi province,one strain of IBV was isolated from chicken.Two pairs of primers for amplifying the N and M genes of IBV were designed according to the sequences in GenBank.The N and M genes of the strain were amplified by RT-PCR,and they were proved to be the N and M genes of IBV by cloning,sequencing and compared with reference IBV strains published in GenBank.The results showed that the N gene from the IBV isolate consisted of 1 230 bp,coding 409 amino acids.The M gene from the IBV isolate consisted of 678 bp,coding 225 amino acids.The sequence analysis of N gene showed that it shared 87.2% to 93.3% nucleotide homologies and 90.0% to 94.4% deduced amino acid sequence homologies with IBV strains from GenBank.The M gene sequence analysis showed that it shared 83.6% to 91.0% nucleotide homologies and 82.7% to 92.9% deduced amino acid sequence homologies.The phylogenetic tree analysis showed that it was closely related to BJ and LX4 strains,and were clustered into one group;But with the distant relatives from other strains of IBV.These results suggested that the isolate was a new variant of IBV.     

Pathogenicity of a Strain of H5N6 Subtype Avian Influenza Virus in Ducks and Mice

To investigate the pathogenicity of A/Hero/Guangdong/C1/2013(H5N6), an AIV strain isolated from Heron on ducks and mice, pathogenicity of this new virus strain and changing of histopathology as well as a preliminary study on its biological characteristics were studied by virus challenges test via intranasal and eye-drop and in chicken via jugular vein injection.Results showed that the EID₅₀ of this strain of virus was 10^-8.16/0.1 mL in embryonated chicken eggs and the intravenous inoculation of pathogenic index (IVPI) was 2.76.In ducks and mice, the 50% lethal doses (LD₅₀) of it were determined to be 10^-4.0/0.2 mL and 10^-4.67/0.05 mL, respectively.Symptoms of infection included loss of appetite, depression, swollen head and tears after being infected with 10⁶ EID₅₀ per duck by intranasal and eye-drop administration.Most of ducks died 4 to 7 days post-infection, liver, lung and kidney still eliminated toxicants at day 7 post-infection.Anatomy showed symptoms of pericardial effusion, pulmonary congestion and kidney enlargement, while pathological section showed pathological change like karyopycnosis and inflammatory cell infiltration in heart, liver, kidney and spleen.Mice developed symptoms of infection like loss of appetite, depression, shaggy coat and ruffled coat after being infected with 5×10⁵ EID₅₀ per mouse by intranasal and eye-drop administration.Most of the infected mice died 5 to 7 days post-infection and only liver still eliminated toxicants at day 7 post-infection.Although organ anatomy showed no obvious pathological changes, pathological section showed pathological changes like karyopycnosis and inflammatory cell infiltration in heart, kidney and lung.Our research demonstrated that this H5N6 subtype AIV had a strong pathogenicity and could be defined as a highly pathogenic AIV strain as its IVPI was greater than 1.2.Our work laid a theory foundation for study, prevention and control of H5N6 subtype AIV.     

Avian Influenza A(H5N1) Virus Outbreak Investigation: Application of the FAO‐OIE‐WHO Four‐way Linking Framework in Indonesia

WHO, FAO and OIE developed a ‘four‐way linking’ framework to enhance the cross‐sectoral sharing of epidemiological and virological information in responding to zoonotic disease outbreaks. In Indonesia, outbreak response challenges include completeness of data shared between human and animal health authorities. The four‐way linking framework (human health laboratory/epidemiology and animal health laboratory/epidemiology) was applied in the investigation of the 193rd human case of avian influenza A(H5N1) virus infection. As recommended by the framework, outbreak investigation and risk assessment findings were shared. On 18 June 2013, a hospital in West Java Province reported a suspect H5N1 case in a 2‐year‐old male. The case was laboratory‐confirmed that evening, and the information was immediately shared with the Ministry of Agriculture. The human health epidemiology/laboratory team investigated the outbreak and conducted an initial risk assessment on 19 June. The likelihood of secondary cases was deemed low as none of the case contacts were sick. By 3 July, no secondary cases associated with the outbreak were identified. The animal health epidemiology/laboratory investigation was conducted on 19–25 June and found that a live bird market visited by the case was positive for H5N1 virus. Once both human and market virus isolates were sequenced, a second risk assessment was conducted jointly by the human health and animal health epidemiology/laboratory teams. This assessment concluded that the likelihood of additional human cases associated with this outbreak was low but that future sporadic human infections could not be ruled out because of challenges in controlling H5N1 virus contamination in markets. Findings from the outbreak investigation and risk assessments were shared with stakeholders at both Ministries. The four‐way linking framework clarified the type of data to be shared. Both human health and animal health teams made ample data available, and there was cooperation to achieve risk assessment objectives.     

黑枣苗氮磷营养及经济施肥的研究

试验采用苗床施肥，结果表明，肥料用量适宜促进营养元素吸收利用，７～８月份苗木叶片含氮量３．５％～４．０％、含磷量０．２％～０．３％、含钾量１．０％或稍高时，苗木生长良好。在该试验条件下，施Ｐ２Ｏ５１２０ｋｇ／ｈａ、施纯氮１８０ｋｇ／ｈａ左右，幼苗产量、质量及施肥利润均达到较高水平。     

赤枝栲干物质生产及氮磷营养动态研究

通过对1年生赤枝栲干物质的生产及氮磷营养动态研究,结果表明:不同的生长季节1年生赤枝栲苗木的各器官生长速率有很大差异,5~7月份根、叶生长迅速,以后迅速下降,8~10月份茎、侧根生长迅速;5~7月份大多数的光合产物用于形成根与叶,少量用于充实茎与侧根的生长;净同化率出现在5~7月份,与根系的生长呈显著的正相关;不同器官在其年生长周期中,氮、磷营养动态表现不同,养分吸收与赤枝栲生长发育规律相吻合。     

核桃叶片中氮、磷、钾含量及年动态变化规律研究

以早实核桃辽宁1号和晚实核桃礼品1号为例，分别测定早实核桃和晚实核桃叶片中不同生育期N、P、K的含量，结果表明，核桃叶片中N、P、K含量在幼龄叶中最高，整个生育期内核桃叶片中N、P、K含量总体上呈逐渐降低。早实核桃的N、P含量比同期的晚实核桃高，且差异极显著；而K含量无差异。     

Nitrogen-fixation dynamics in a cut-and-carry silvopastoral system in the subhumid conditions of Guadeloupe, French Antilles

This paper summarizes several studies on N recycling in a tropical silvopastoral system for assessing the ability of the system to increase soil fertility and insure sustainability. We analyzed the N₂ fixation pattern of the woody legume component (Gliricidia sepium), estimated the recycling rate of the fixed N in the soil, and measured N outputs in tree pruning and cut grass (Dichanthium aristatum). With this information, we estimated the N balance of the silvopastoral system at the plot scale. The studies were conducted in an 11-year-old silvopastoral plot established by planting G. sepium cuttings at 0.3 m × 2 m spacing in natural grassland. The plot was managed as a cut-and-carry system where all the tree pruning residues (every 2-4 months) and cut grass (every 40-50 days) were removed and animals were excluded. No N fertilizer was applied. Dinitrogen fixation, as estimated by the ¹⁵N natural abundance method, ranged from 60-90% of the total N in aboveground tree biomass depending on season. On average, 76% of the N exports from the plot in tree pruning (194 kg [N] ha^–1 yr^–1) originated from N₂ fixation. Grass production averaged 13 Mg ha^–1 yr^–1 and N export in cut grass was 195 kg [N] ha^–1 yr^–1. The total N fixed by G. sepium, as estimated from the tree and grass N exports and the increase in soil N content, was about 555 kg [N] ha^–1 yr^–1. Carbon sequestration averaged 1.9 Mg [C] ha^–1 yr^–1 and soil organic N in the 0-0.2 m layer increased at a rate of 166 kg [N] ha^–1 yr^–1, corresponding to 30% of N₂ fixation by the tree. Nitrogen released in nodule turnover (10 kg [N] ha^–1 yr^–1) and litter decomposition (40 kg [N] ha^–1 yr^–1) contributed slightly to this increase, and most of the recycled N came from the turnover or the activity of other below-ground tree biomass than nodules. This revised version was published online in June 2006 with corrections to the Cover Date.     

One-day rate measurements for estimating net nitrification potential in humid forest soils

Measurements of net nitrification rates in forest soils have usually been performed by extended sample incubation (2–8 weeks), either in the field or in the lab. Because of disturbance effects, these measurements are only estimates of nitrification potential and shorter incubations may suffice. In three separate studies of northeastern USA forest soil surface horizons, we found that laboratory nitrification rates measured over 1 day related well to those measured over 4 weeks. Soil samples of Oa or A horizons were mixed by hand and the initial extraction of subsamples, using 2 mol L⁻¹ KCl, occurred in the field as soon as feasible after sampling. Soils were kept near field temperature and subsampled again the following day in the laboratory. Rates measured by this method were about three times higher than the 4-week rates. Variability in measured rates was similar over either incubation period. Because NO₃⁻ concentrations were usually quite low in the field, average rates from 10 research watersheds could be estimated with only a single, 1-day extraction. Methodological studies showed that the concentration of NH₄⁺ increased slowly during contact time with the KCl extractant and, thus, this contact time should be kept similar during the procedure. This method allows a large number of samples to be rapidly assessed.