特殊医学用途婴儿配方食品中氯的国标检测方法适用性

依据GB 5009.44—2016《食品安全国家标准 食品中氯化物的测定》，对特殊医学用途婴儿配方食品中氯的检测方法适用性展开研究。结果表明：采用GB 5009.44—2016第三法（银量法）检测特殊医学用途婴儿配方食品中乳蛋白部分水解配方、乳蛋白深度水解配方、氨基酸配方得到的样品加标回收率分别为95%～108%、98%～125%、100%～106%，相对标准偏差为12.7%～32.9%；采用GB 5009.44—2016第一法（电位滴定法）得到的检测结果低于第三法检测结果。     

盐胁迫对向日葵幼苗生长及生理特性的影响

以银叶向日葵ARG1807（Wt）为供试材料，陕葵2号的保持系（HA）和恢复系（HL）为对照材料，利用250 mmol·L^-1 NaCl溶液进行盐胁迫处理7 d，分析供试材料与对照材料盐胁迫前后的形态变化以及叶片中的叶绿素、脯氨酸、丙二醛含量及SOD活性等生理指标差异，探究银叶向日葵耐盐的形态和生理基础。结果表明：（1）Wt的株高、叶面积增量以及根、茎、叶生物量分别比未进行NaCl胁迫处理植株（对照，CK）减少24.53%、38.21%、2.59%、5.50%、3.42%，其变化幅度小于HA和HL。(2) 在盐胁迫处理下，各向日葵品种的胞间二氧化碳浓度（Ci）均显著下降，而气孔限制值（Ls）显著上升，说明HA、HL与Wt净光合速率降低主要是由于气孔因素。（3）在盐胁迫下，3个向日葵品种叶片的脯氨酸含量（Pro）、丙二醛含量（MDA）、超氧化物歧化酶（SOD）活性、过氧化物酶（POD）活性以及过氧化氢酶活性（CAT）均发生了显著变化，其中，Wt的脯氨酸含量、SOD活性、POD活性以及CAT活性分别比CK增加279.94%、65.09%、211.11%和55.91%，高于HA 和HL，而MDA含量比CK增加231.10%，低于HA。（4）随着时间的推移，3个向日葵品种叶片失水速率均显著上升，其中，Wt的失水速率一直低于HA、HL。在失水150 min时，HA失水速率最高，达84%，Wt叶片失水最慢，失水速率为69%，说明Wt在保水能力方面具有显著优势。（5）观察向日葵叶片表型发现：Wt的叶片表皮毛长度长，密度明显多于HA和HL，推测这可能是Wt保水性强于HA和HL的原因。分析表明：在盐逆境下，形态指标和生理指标，均为银叶向日葵表现最佳，优势突出，表明其在盐逆境下具有更强的适应能力。     

Assessment of the effect of silver nanoparticles on the relevant soil protozoan genus Acanthamoeba

Silver nanoparticles (AgNP) are used in a broad range of consumer products and industrial applications. During the regular product life cycle and disposal, AgNP are continuously released into the environment. Hence, the aim of this study was to investigate the potential ecotoxicological effects of AgNP exposure on amoebae. The Acanthamoeba castellanii ATCC 30234 strain and environmental isolate Acanthamoeba strain C5/2, which are both affiliated with genotype T4, were chosen as representatives of ecologically important soil protozoan organisms. The amoebae were exposed to citrate‐stabilized AgNP (30 and 70 nm in size) for 24 h and 96 h at concentrations ranging from 600 µg L⁻¹ to 20 mg L⁻¹. A newly adopted cell culture based microscopic assay was applied to assess the adherence ability of the amoeba trophozoites. The general metabolic activity of Acanthamoeba was determined to be a second independent endpoint by means of intracellular reduction of the redox dye AlamarBlue®. The fate of AgNP within the amoebae and test solutions was visualized by light‐ and transmission electron microscopy (TEM). Both Acanthamoeba strains showed a significant dose‐dependent decrease of adherence ability (p  < 0.04) and metabolic activity (p < 0.01) after 96 h of AgNP exposure. The environmental Acanthamoeba strain C5/2 lost both its adherence ability and metabolic activity at lower AgNP concentrations than the type strain, indicating a higher sensitivity to ionic silver. This was confirmed by the application of AgNO₃, provoking a higher effect level in strain C5/2. AgNP was visualized intracellularly by transmission electron microscopy within the cytoplasm of Acanthamoeba . This is the first report to show the ecotoxicological effects of short‐term AgNP exposure on the soil protist Acanthamoeba , causing both changes in the adherence ability and metabolic activity of this amoeba. This combined approach may be a powerful tool in the future for predicting potential harmful ecotoxicological effects of AgNP exposure using soil protozoans.     

黑褐新糠虾血细胞形态及其与日本新糠虾对溶藻弧菌敏感性的比较

实验采用Giemsa与Wright' s两种染色方法对黑褐新糠虾血细胞进行染色,根据血细胞大小,所含颗粒与否,颗粒多少和核质比将其分型,并对各型血细胞所占比例进行统计.结果表明,通过比较Giemsa染色与Wright's染色两种方法的染色时间和效果,Wright's染色更加适合糠虾血细胞.通过染色,将黑褐新糠虾血细胞分为3种类型,它们分别为透明细胞、半颗粒细胞和颗粒细胞.它们的胞体大小依次增大,核质比依次降低.对此3种类型血细胞比例分别统计后发现,颗粒细胞所占比例最多约( 48.38％±0.05％),其次是半颗粒细胞约为(35.22％±0.04％),最少的为透明细胞约为(16.40％±0.01％).对不同浓度溶藻弧菌的感染试验发现,日本新糠虾在较短的时间内出现死亡,即48h,而黑褐新糠虾72 h出现死亡,各浓度组中日本新糠虾死亡率均高于黑褐新糠虾.黑褐新糠虾相对日本新糠虾来说,对溶藻弧菌具有较低的敏感性,可能与其血细胞中颗粒和半颗粒比例较高有关.     

VEGF在绵羊生殖管道中的表达规律

以不同发情周期雌性绵羊子宫、输卵管为研究对象,采用免疫组织化学技术,针对血管内皮生长因子（VEGF）在绵羊子宫、输卵管的表达、定位和变化规律进行了检测,同时应用相关图像分析软件对抗原染色强度进行了定量分析。结果表明：输卵管在发情0～15d,VEGF表达量在第9天达到峰值后经历波动逐渐下降过程,输卵管内膜上皮细胞是VEGF抗原的主要靶细胞;而子宫角在发情0～15d,VEGF表达量在第5天达到峰值后经历波动逐渐下降过程,子宫内膜固有层及腺体周围细胞为VEGF抗原的主要靶细胞。该研究结果为绵羊生产中进一步提高受胎率和妊娠率及频密产羔等技术的应用提供了科学依据。     

花生种子特异启动子AHSSP1的克隆及功能分析

为丰富花生种子特异启动子资源,本研究利用PCR技术在花生基因组中克隆了种子贮藏蛋白基因PSC32的启动子AHSSP1,利用半定量RT-PCR检测了PSC32基因表达模式,借助NewPLACE在线分析了AHSSP1序列中存在的顺式作用元件,并构建了AHSSP1驱动GUS报告基因的表达载体,经农杆菌转化获得转基因拟南芥,经GUS组织化学染色鉴定了该启动子的功能。结果表明,PSC32基因957 bp长的启动子AHSSP1序列具备种子特异表达启动子特有的3个RY REPEAT元件。半定量RT-PCR分析发现,PSC32基因在花生成熟种子中表达,而在饱果成熟期根、茎、叶片、花、入土前的果针、成熟种子的果壳中均不表达。GUS组织化学染色发现,转基因拟南芥成熟种子以及萌发种子的子叶、下胚轴和胚根均能够被染上蓝色;长出真叶后,子叶和下胚轴仍能被染色,而根和真叶不能被染上蓝色;成年期转基因拟南芥的叶片也不能被染上蓝色。而野生型拟南芥整个生长时期均不能被染上蓝色。以上现象说明AHSSP1是一个种子特异启动子。本研究丰富了花生种子特异启动子的资源,对花生籽仁品质改良或以花生籽仁作为“生物反应器”的研究具有重要的应用价值。     

Effect of dietary graded levels of cottonseed meal and gossypol on growth performance,body composition and health aspects of allogynogenetic silver crucian carp,Carassius auratus gibelio♀ × Cyprinus carpio♂

A 6‐month trial was conducted to evaluate the effects of dietary cottonseed meal (CSM) and free gossypol (FG) on allogynogenetic silver crucian carp, Carassius auratus gibelio♀ × Cyprinus carpio♂ with 4 replicates of each treatment. Isonitrogenous and isocaloric diets were formulated with the 0 g kg^?1 (control), 200 g kg^?1, 400 g kg^?1, and 560 g kg^?1 CSM. Diets with FG were made by supplementing batches of control diet with 214 mg kg^?1, 428 mg kg^?1, and 642 mg kg^?1. Weight gain, specific growth rate, and protein efficiency ratio increased significantly up to an inclusion level of CSM of 400 g kg^?1 in the diet, with a significant decrease in food conversion ratio. Further increase in CSM to 560 g kg^?1 did not cause further changes in fish performance. Free gossypol did not affect fish performance significantly at any inclusion level. Neither CSM nor FG caused significant effects in any of the other evaluated parameters such as whole body composition, haemoglobin concentration, activities of serum lysozyme, superoxide dismutase, alanine aminotransferase and aspartate aminotransferase, and histology of hepatic tissues and midgut. Our results suggested that crucian carp can tolerate at least 642 mg kg^?1 FG and that it is safe to including 400 g kg^?1 CSM in crucian carp feed.     

Essential oil of Aloysia citriodora Paláu and citral: sedative and anesthetic efficacy and safety in Rhamdia quelen and Ctenopharyngodon idella

ObjectiveTo verify the efficacy of citral in inducing sedation and anesthesia in silver catfish (Rhamdia quelen) and grass carp (Ctenopharyngodon idella) and to assess the safety of essential oil (EO) of Aloysia citriodora and citral in inducing and maintaining anesthesia in silver catfish.Study designClinical study, randomized, parallel, multi-arm with control group in target species.AnimalsA total of 96 juvenile and 72 adult silver catfish and 80 juvenile grass carp were used.MethodsSilver catfish and grass carp were exposed to different concentrations of citral, 15–675 and 15–600 μL L^–1, respectively, during the maximum period of 30 minutes to verify sedation and anesthesia induction and recovery times. In addition, for anesthetic induction, silver catfish were exposed to the EO of A. citriodora and citral at 225 μL L^–1 for 3.5 minutes. Then, fish were transferred to an anesthesia maintenance solution at 50 μL L^–1 for 10 minutes to assess hematologic and biochemical variables at 60 minutes, 2 and 6 days after treatment.ResultsCitral only induced sedation from 15, 25 and 40 μLL^–1 in both species. Anesthesia without mortality was induced in silver catfish at 50–600 μL L^–1 and grass carp at 75–450 μL L^–1. At 675 and 600 μL L^–1, mortality was recorded in silver catfish and grass carp, respectively. The EO of A. citriodora and citral were safe in inducing and maintaining anesthesia in silver catfish, with mean corpuscular hemoglobin concentration being the only variable that varied in relation to time and treatments.Conclusions and clinical relevanceCitral was effective in inducing sedation and anesthesia in both species. In addition, A. citriodora EO and citral were safe in inducing and maintaining anesthesia in silver catfish. Both agents are promising substances for the development of new drugs for fish.     

纳米银体外抗猪繁殖与呼吸综合征病毒的作用研究

【目的】 研究纳米银（silver nanoparticles,AgNPs）体外抗猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus,PRRSV）的作用,并初步分析其抗病毒作用机制,为PRRSV的防控提供新思路。【方法】 使用0.1875、0.375、0.75、1.5、3、6、12 μg/mL AgNPs处理Marc145细胞,采用CCK-8试剂盒评估AgNPs对Marc145细胞毒性,确定其安全浓度。通过显微观察、间接免疫荧光试验、病毒滴度测定、实时荧光定量RT-PCR方法评估AgNPs体外抗PRRSV感染Marc145细胞的效果。通过间接免疫荧光试验和实时荧光定量RT-PCR方法评估AgNPs对PRRSV的直接灭活效果。通过实时荧光定量RT-PCR方法分析AgNPs对不同感染复数（multiplicity of infection,MOI） PRRSV （0.0001~0.1）黏附和入侵Marc145细胞的影响,以及PRRSV感染Marc145细胞3、6、12、18和24 h后加入AgNPs对Marc145细胞增殖的影响。【结果】 AgNPs对Marc145细胞的最大安全浓度为1.5 μg/mL,0.375、0.75、1.5 μg/mL AgNPs均具有良好的体外抗PRRSV活性,0.375、0.75、1.5 μg/mL AgNPs均对PRRSV起一定灭活作用。AgNPs对不同MOI的毒株黏附和入侵Marc145细胞均有一定抑制作用,且不同时间（3、6、12、18、24 h）加入AgNPs对PRRSV增殖均有一定抑制作用。【结论】 AgNPs具有良好的体外抗PRRSV活性,体外抗PRRSV的作用机制包括直接灭活以及抑制病毒的黏附、入侵和增殖过程。     

A rapid differential staining technique for Fusarium pseudograminearum in cereal tissues during crown rot infections

A novel fluorescence‐based differential staining procedure has been developed for the rapid visualization of Fusarium pseudograminearum hyphae in cereal tissues. Infected tissues are stained with safranin, which binds to host cell walls, while hyphal tissues are stained with the fluorescent dye solophenyl flavine 7GFE. The method is rapid, does not require clearing of culm cross‐sections, and provides high contrast informative images of fungal and plant structures during pathogenesis.