柑橘轮斑病的适生区预测及风险分析

【目的】 柑橘轮斑病（citrus target spot）作为一种新发柑橘病害,造成发病果园严重的经济损失。本研究针对该病害进行适生区预测及风险分析,以便对该病采取及时、有效的管控措施,最终达到降低其流行风险等级,防止病害传播扩展的目的。【方法】 基于环境变量数据和柑橘轮斑病发生分布数据,运用MaxEnt生态位模型模拟预测柑橘轮斑病菌（Pseudofabraea citricarpa）在中国的潜在适生区分布。并通过ROC（receiver operating characteristic）曲线下面积（area under the curve,AUC）评估预测模型的精度,运用正规化训练增益刀切法（regularized training gain）获取气候因子与分布概率间的关系。同时采用有害生物风险分析理论,以有害生物风险分析的规定程序为依据探索柑橘轮斑病病害的风险分析体系和评价值的计算方法,对评价指标进行定性分析,进而量化评价值。在建立综合评价模型的基础上,计算柑橘轮斑病风险性危害值,最后对病害的风险性危害值进行评价。【结果】 柑橘轮斑病菌MaxEnt模型预测结果的平均AUC值为0.998,表明预测结果精度高。柑橘轮斑病菌的潜在适生区面积约占全国面积12.19%,高适生区、中适生区、低适生区各占全国面积约2.85%、3.99%、5.35%。高、中适生区主要集中于长江中上游柑橘优势区及其周边。其中,高适生区主要集中在四川、重庆、陕西南部,以及贵州、湖北等少量地区。中、低适生区是高适生区的外围扩展。通过MaxEnt模型正规化训练增益刀切法获取的环境变量重要性分析结果表明,最冷季度平均温度（Bio11）、最干季度平均温度（Bio9）、最冷月最低温（Bio6）是影响柑橘轮斑病菌分布的3个关键环境因子,这意味着低温、干冷季节柑橘轮斑病发生可能性大。风险分析最终创建出5个准则层、13个指标层的多指标综合评价体系,并对各指标层定量与定性分析,柑橘轮斑病在我国的风险性危害值（R值）为2.08,处于高度风险等级,对长江中上游及湖北西部-湖南西部两大柑橘产区的潜在危害最大。【结论】 柑橘轮斑病风险性较高,需要尽快建立监测体系,针对病害采取有效控制措施,阻止病害在长江中上游柑橘优势区及相邻柑橘产区传播。     

Bilateral apex pubis distance：a novel index for follicular development and egg laying status in domestic hens (Gallus gallus domesticus)

ABSTRACT

1. Measures of the bilateral apex pubis (BAP) in White Leghorn (WLH) hens aged from 16 to 64 weeks (from the time ovaries were completely undeveloped to senility) were ranked into various groups so the relationship between external traits (BAP distance, comb length, comb height, wattle length, wattle height, body weight) and internal traits (largest ovarian follicle diameter (LFD) and oviduct weight and length) could be determined.

2. Using correlation and regression analysis with data from 16 weeks of age to the onset of lay, measurements of the apex pubis showed a strong correlation with LFD (r = 0.80, P < 0.01) and oviduct weight (r = 0.80, P < 0.01).

3. Multiple comparisons among WLH hens from 16 to 43 weeks of age showed that the BAP distance increased in all groups ranked by LFD or by laying period.

4. In the late laying period (>60 weeks of age), BAP distance showed a significant decrease (P < 0.01) in long-term non-laying hens compared to laying hens.

5. These findings suggest BAP distance can serve as an index for evaluating the onset and probable stages of follicular development during sexual maturation and/or aid in the identification and selection of non-laying hens during reproduction.     

First serological evidence of West Nile virus infection in wild birds in Northern Algeria

While the epidemiology of Flaviviruses has been extensively studied in most of the Mediterranean basin, little is known about the current situation in Algeria. In order to detect the circulation of West Nile (WNV) and Usutu viruses (USUV) in Kabylia, 165 sera were collected from two wild birds species, namely the long distance migrant Turdus philomelos (song thrush) (n = 92) and the resident Passer domesticus (house sparrow) (n = 73). A total of 154 sera were first analyzed by commercial competition ELISA. WNV and USUV micro-neutralization tests were performed on all c-ELISA positive sera and all samples with poor volume. Overall, 7.8 % (CI95 %: 3.5–11.9) were positive by c-ELISA. Positive results were detected in 12.5 % (CI95 %:5.6–19.4) of song thrushes and 1.5 % (CI95 %: 0.0–4.5) for sparrow.Micro-neutralization tests revealed an overall seroprevalence of 6.7 % for WNV (CI95 %: 2.9–10.3), Neutralizing antibodies were found in 8.7 % (CI95 %: 3.0–14.4) for song thrushes and in 4.1 % (CI95 %: 0.0–8.7) of sparrows. The current study demonstrates significant seroprevalence of WNV antibodies in wild birds in Algeria.     

Ethyl acetate fraction of flavonoids from Polygonum hydropiper L. modulates pseudorabies virus-induced inflammation in RAW264.7 cells via the nuclear factor-kappa B and mitogen-activated protein kinase pathways

Pseudorabies virus (PRV) infection leads to severe inflammatory responses and tissue damage, and many natural herbs exhibit protective effects against viral infection by modulating the inflammatory response. An ethyl acetate fraction of flavonoids from Polygonum hydropiper L. (FEA) was prepared through ethanol extraction and ethyl acetate fractional extraction. An inflammatory model was established in RAW264.7 cells with PRV infection to evaluate the anti-inflammatory activity of FEA by measuring cell viability, nitric oxide (NO) production, reactive oxygen species (ROS) release, and mRNA expression of inflammatory factors, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Its functional mechanism was investigated by analyzing the phosphorylation and nuclear translocation of key proteins in the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Our findings indicate that PRV induced inflammatory responses in RAW264.7 cells, and the responses were similar to that in lipopolysaccharide (LPS)-stimulated cells. FEA significantly suppressed NO synthesis and down-regulated both expression and secretion of COX-2, iNOS, and inflammatory cytokines (P<0.05 or P<0.01). FEA also reduced NF-κB p65 translocation into the nucleus and decreased MAPK phosphorylation, indicating that the NF-κB/MAPK signaling pathway may be closely related to the inflammatory response during viral infection. The findings suggested the potential pharmaceutical application of FEA as a natural product that can treat viral infections due to its ability to mitigate inflammatory responses.     

链霉菌30702复合壳聚糖对番木瓜PRSV的防控效果

探究链霉菌30702复合壳聚糖对番木瓜环斑病毒病（papaya ringspot virus，PRSV）的防控效果，为番木瓜环斑花叶病防治提供基础。以台农二号番木瓜为材料，壳聚糖设3个浓度1.0g/L、0.5g/L、0.25g/L；30702菌液2个浓度0 cfu/L和4.2×10 6cfu/L，互相组合为6个处理，以清水处理为空白组，宁南霉素0.05 g/L为对照组，共8个处理；运用分光光度计法测量相关酶活。1.0 g/L壳聚糖复合4.2×106 cfu/L链霉菌30702的防治效果为88.6（p＜0.05），显著高于其他组；壳聚糖复合链霉菌30702组的防治效果、超氧化物歧化酶(SOD)活性、过氧化物酶（POD）活性、多酚氧化酶(PPO)活性相较于单独使用壳聚糖组有显著提升。使用1.0 g/L壳聚糖复合4.2×106 cfu/L的30702对番木瓜防控PRSV的效果最好，防治指数达到88.6%。     

猪皮炎肾病综合征诊断与防治

猪皮炎肾病综合症是由圆环病毒二型感染引起的一种以猪皮肤表面出现红疹、丘疹、结痂或坏死,肾脏严重肿大为主要临床特征的常见传染性疾病。皮炎肾病综合症主要危害生长猪和育肥猪。受病毒感染后,患病猪表现为生长发育不良,免疫功能受到严重抑制,易继发感染其他传染性疾病,表现出复杂的临床症状,给生猪养殖业造成严重损失和危害。防范疾病的发生流行,需要掌握猪皮炎肾病综合症的具体流行特点、临床表现,并采取针对有效措施进行诊断,然后构建有效的防控措施,短时间内控制病情,避免损失进一步加大。该文主要论述猪皮炎肾病综合症的流行特点、临床症状、病理变化、诊断方法和防治措施。     

柑橘脉突病毒基因组全长cDNA克隆及其侵染性鉴定

【目的】构建柑橘脉突病毒（citrus vein enation virus,CVEV）侵染性克隆,为从分子水平解析其致病机理打下基础。【方法】利用SMARTer^? RACE(rapid amplification of cDNA ends)试剂盒对CVEV的5′序列进行RACE,并依据序列分析结果及CVEV分离株VE-1保守序列,设计CVEV基因组全长cDNA扩增引物。以CVEV毒源植株的总RNA为模板,通过EV25-F/EV5983-R引物扩增CVEV基因组全长cDNA。利用In-Fusion重组连接线性化pXT1和CVEV全长cDNA。通过菌液PCR及测序分析鉴定CVEV基因组全长cDNA克隆。通过农杆菌介导的真空浸润接种摩洛哥酸橙（Citrus aurantium）、邓肯葡萄柚（C. paradisi）、尤力克柠檬（C.limon）、枳柚（C. paradisi×Poncirus trifoliata)、Rusk枳橙（P. trifoliata×C. sinensis）、枣阳小叶枳(P. trifoliata),进一步通过RT-PCR检测、症状观察鉴定所构建CVEV全长cDNA克隆的侵染性。【结果】建立了CVEV的基因组全长RT-PCR扩增体系,获得基于双元载体pXT1的CVEV基因组全长cDNA克隆10个。随机选取的6个全长cDNA克隆CVEV1901—CVEV1906的序列一致性为99.35%。其中,CVEV1901基因组全长5 983 nt,由5个开放阅读框、5′端207 nt和3′端198 nt的两个非翻译区、以及ORF2和ORF3之间122 nt的基因间隔区组成。序列分析结果显示,CVEV1901与浙江分离株XZG及四川SM分离株的序列一致性分别为99.98%和99.11%;与西班牙VE-1分离株、美国加州VE701分离株和日本IBK分离株基因组序列一致性在96.89%—98.61%;与同属中豌豆耳突花叶病毒（pea enation mosaic virus）和紫花苜蓿耳突病毒（alfalfa enamovirus）的序列一致性约90%。通过农杆菌介导的真空浸润将CVEV1901接种至6个不同的柑橘品种,接种后120 d的RT-PCR检测结果表明摩洛哥酸橙、邓肯葡萄柚、尤力克柠檬、枳柚、Rusk枳橙和枣阳小叶枳阳性植株/接种植株（阳性率）分别为16/17（94.12%）、12/14（85.71%）、16/21（76.19%）、15/19（78.95%）、13/14（92.86%）和0/18（0）。其中,部分摩洛哥酸橙出现典型CVEV侵染症状,叶片侧脉和支脉产生耳状小突起,叶背有相应的凹陷;部分邓肯葡萄柚和尤力克柠檬出现叶片皱缩现象。【结论】建立了CVEV的基因组全长RT-PCR扩增体系,获得了CVEV基因组全长cDNA侵染性克隆,通过农杆菌介导的真空浸润接种可引起摩洛哥酸橙、邓肯葡萄柚和尤力克柠檬的CVEV侵染症状。     

甜椒L 3应对辣椒轻斑驳病毒及中椒系列新品种的选育

辣（甜）椒是我国栽培面积最大的蔬菜作物,年播种面积约2.2×10⁶ hm²,其中甜椒约5×10⁵hm²。生产上传统病害如疫病、病毒病等依然严峻,近年来辣椒轻斑驳病毒（PMMoV,烟草花叶病毒属）等新型流行病害爆发,严重制约甜椒生产;同时,消费者对品种的品质、多样性提出更高要求。笔者课题组先后从国内外引进甜（辣）椒种质资源1 400余份,通过鉴定、评价,筛选出具有抗病毒病、白粉病和果大、皮薄、光泽度好、品质优等优良性状的种质资源120余份。构建了与抗TMV（L³、L⁴）、抗番茄斑点萎蔫病毒TSWV（Tsw）等重要性状紧密连锁的分子标记,辅助育种准确率达90%以上。通过常规育种技术和分子标记相结合,创制出含有抗PMMoV（P_0,1,2）L³,且兼具抗TMV、ToMV、PMMoV（P_0,1,2）、CMV和疫病,果大、果实均一度高、光泽度好等综合性状优良、配合力高的甜椒骨干亲本‘0516’,以其为骨干亲本,培育出4个新一代优质、多抗、适应不同生态区的新品种‘中椒105号’‘中椒106号’‘中椒107号’‘中椒108号’。上述系列新品种含有L ³,抗TMV、PMMoV（P_0,1,2),兼抗CMV和疫病;果实形状大小、色泽、整齐度等商品品质,Vc等营养品质显著提高。     

A Lymphoid Organ Specific Anti-Lipopolysaccharide Factor from Litopenaeus vannamei Exhibits Strong Antimicrobial Activities

Different shrimp species are known to possess apparent distinct resistance to different pathogens in aquaculture. However, the molecular mechanism underlying this finding still remains unknown. One kind of important antimicrobial peptides, anti-lipopolysaccharide factors (ALF), exhibit broad-spectrum antimicrobial activities. Here, we reported a newly identified ALF from the shrimp Litopenaeus vannamei and compared the immune function with its counterpart in the shrimp Fenneropenaeus chinensis. The ALF, designated as LvALF8, was specifically expressed in the lymphoid organ of L. vannamei. The expression level of LvALF8 was apparently changed after white spot syndrome virus (WSSV) or Vibrio parahaemolyticus challenges. The synthetic LBD peptide of LvALF8 (LvALF8-LBD) showed strong antibacterial activities against most tested Gram-negative and Gram-positive bacteria. LvALF8-LBD could also inhibit the in vivo propagation of WSSV similar as FcALF8-LBD, the LBD of LvALF8 counterpart in F. chinensis. However, LvALF8-LBD and FcALF8-LBD exhibited apparently different antibacterial activity against V. parahaemolyticus, the main pathogen causing acute hepatopancreatic necrosis disease (AHPND) of affected shrimp. A structural analysis showed that the positive net charge and amphipathicity characteristics of LvALF8-LBD peptide were speculated as two important components for its enhanced antimicrobial activity compared to those of FcALF8-LBD. These new findings may not only provide some evidence to explain the distinct disease resistance among different shrimp species, but also lay out new research ground for the testing and development of LBD-originated antimicrobial peptides to control of shrimp diseases.     

瓜类褪绿黄化病毒及其系统进化分析

【目的】检测新疆南疆7个县/市瓜类褪绿黄化病毒(Cucurbit chlorotic yellows virus, CCYV),进行系统发育分析,为新疆南疆CCYV预警和防控提供参考。【方法】于2019年,从中国新疆南疆7个县/市采集带有黄化特征的51份甜瓜叶片样品,RT-PCR进行CCYV的检测,进行特异性片段的克隆、测序和系统发育分析。【结果】中国新疆巴楚县、阿克苏市、莎车县、伽师县、疏勒县、疏附县均未检测到CCYV,而洛浦县的13份样本中,8份检出为CCYV阳性,检出率为61.53%,洛浦县CCYV特异性片段克隆测序获得了685 bp的核酸序列,与GenBank中的CCYV分离物外壳蛋白(Coat protein, CP)的一致性达到100%。所得序列与中国新疆(吐鲁番)、中国其他省、苏丹、日本、塞浦路斯、黎巴嫩的CP基因聚在I组(Group),沙特阿拉伯的分离物聚在Ⅱ组,伊朗的分离物聚在Ⅲ组。【结论】CCYV在中国新疆南疆洛浦县甜瓜产区发生,与中国新疆吐鲁番、中国其他省及周围国家的CCYV分离物亲缘关系很近,且群体遗传变异很小。