水族馆养殖中华鲟血细胞指标的变化

血细胞是临床健康的重要指标。本文对北京海洋馆养殖的40尾中华鲟（Acipenser sinensis）外周血细胞组成进行研究,研究对象包括超过30龄、18龄、16龄、15龄、11龄、7龄和4龄共7个龄组,并分析比较不同年龄、性腺快速发育前后以及处于疾病前后血细胞指标变化,希望应用于健康评价。结果表明,1）外周血红细胞总数（RBC）平均为（5.56 ± 1.19）× 1011/L,18龄和11龄与其他龄组之间存在显著性差异（P < 0.05）,超过30龄组血细胞比积（HCT）和血红蛋白（HB）值显著高于其他龄组（P < 0.05）;2）白细胞总数（WBC）平均为（16.53 ± 4.94）× 109/L,18龄与4龄间存在显著差异,且分别与其他龄组间存在显著性差异（P < 0.05）。3）白细胞分类计数（DLC）中各类型白细胞平均所占百分比分别为,大淋巴细胞（LLYM）5.26% ± 3.95%、小淋巴细胞（SLYM）77.74% ± 11.73%、嗜中性粒细胞（NEU）9.40% ± 7.98%、嗜酸性粒细胞（EOS）1.90% ± 2.06%和单核细胞（MON）5.50% ± 4.00%,超过30龄和4龄间显著差异,且分别与其他龄组间存在显著性差异（P < 0.05）。4）性腺进入快速发育阶段后,红细胞总数、血细胞比积和白细胞总数较发育前均显著增加（P < 0.05）,血红蛋白值有增加但差异不显著。5）疾病中华鲟白细胞总数显著增高（P < 0.05）并伴随红细胞各指标降低,白细胞数增加幅度达90%,分类计数中中性粒细胞和单核细胞比例显著变化（P < 0.05）。中华鲟快速发育阶段及疾病状态的血细胞组成不同,血细胞密度的变化以及分类计数的差异可及时判断中华鲟健康状况,结果对中华鲟的健康评价与保育研究有重要的参考意义。     

幼虫密度对草地螟血细胞数量和组成的影响

为了阐明幼虫密度对草地螟Loxostege sticticalis L.（鳞翅目： 螟蛾科）细胞免疫能力的影响, 本研究调查了在活体灰菜植株上1,5,10和20头/瓶（900 mL）4种密度条件下的其5龄幼虫血细胞种类、数量和组成。结果表明： 草地螟幼虫血淋巴中有原血细胞、浆血细胞、 颗粒血细胞、珠血细胞和类绛色血细胞等5种（类）血细胞。血细胞总数、 浆血细胞、颗粒血细胞数量随幼虫密度的增加而显著递增, 但原血细胞、珠血细胞和类绛色血细胞数量在幼虫密度间的差异不明显;各种血细胞所占血细胞总数的比例在4个密度中的排序相同, 但10和20头/瓶密度下的浆血细胞比例显著高于1头/瓶的,其余4种血细胞的比例在不同密度之间无显著差异。可见, 幼虫密度主要是通过影响草地螟幼虫浆血细胞和颗粒血细胞的数量及血细胞总数, 从而影响草地螟的细胞免疫能力。     

家蚕幼虫血细胞密度变化成因及血细胞密度与耐高温性的关系

[目的]血细胞是昆虫血淋巴免疫的主导者.调查家蚕Bombyx mori幼虫血细胞密度变化和成因、血细胞密度与家蚕抗性的关系,是研究家蚕血细胞相关的免疫调控和抗性育种的重要组成.[方法]用细胞计数板统计家蚕品种大造不同龄期(4龄第1-4天、5龄第1-8天和上蔟期)幼虫10 μL血淋巴中的血细胞数目并计算血细胞密度,利用I...     

嘴壶夜蛾的形态、生活史及昼夜节律

嘴壶夜蛾Oraesia emarginata (Fabricius)是危害水果果实的重要害虫之一, 其成虫和幼虫取食不同的寄主植物, 可以作为理想的嗅觉研究模式昆虫。为了全面地了解嘴壶夜蛾的形态特性和生物学特性, 本实验通过室内饲喂和红外摄像机观察, 对嘴壶夜蛾各虫态的外部形态、 发育以及昼夜活动节律进行了系统研究。结果表明： 嘴壶夜蛾的各龄幼虫可以通过体色, 体表色斑的类型、 位置和数量, 以及腹足的数量进行区别。通过蛹的生殖孔和成虫触角能够很好地区别雌雄。在室内饲养条件下, 嘴壶夜蛾的寿命为53.18±1.70 d, 存活率为63.62%±2.15%, 其中幼虫的发育历期最长, 存活率最低, 卵的发育历期最短, 存活率最高, 雄成虫的存活时间显著长于雌成虫（P=0.008）。6龄幼虫的发育历期（5.29±0.15 d）显著长于其余各龄幼虫（P<0.001）。同一龄发育中期幼虫的体长和体重显著大于将蜕皮幼虫和刚蜕皮幼虫（P≤0.037）; 第2-6龄刚蜕皮幼虫的体重和体长与前一龄发育中期幼虫之间没有显著差异（P≥0.106）。幼虫在光期的孵化、 蜕皮和化蛹比例高于暗期, 而成虫在暗期的飞行、 产卵比例高于光期, 成虫的飞行随着暗期时间的增加而逐渐变少, 光期成虫的飞行主要在开灯之后1 h。成虫交配集中在暗期的第3-5 小时。本研究结果有助于制定有效的嘴壶夜蛾防治措施, 而且为嘴壶夜蛾作为嗅觉研究模式奠定基础。     

瓜实蝇耐低温相关生理指标的测定

【目的】了解瓜实蝇Bactrocera cucurbitae(Coquillett)对低温的耐受性,分析其不同发育阶段耐寒力差异。【方法】以不同发育阶段瓜实蝇为实验材料,分别测定并比较了过冷却点、水分、脂肪、甘油和糖类含量的变化情况。【结果】其不同龄期的过冷却点差异显著,5日龄蛹的过冷却点最低,为﹣17.04℃,1日龄幼虫的过冷却点最高,为﹣11.82℃,同一日龄的雌、雄成虫之间的过冷却点无显著差异;其幼虫期和蛹期虫体的含水量随着龄期的增长而下降,从1日龄幼虫79.64%下降到5日龄蛹65.31%,5日龄蛹的含水量显著低于其他龄期瓜实蝇,成虫期各龄期雌成虫含水量均显著高于雄成虫;5日龄幼虫脂肪含量最高,为32.90%,1日龄雄成虫和14日龄雄成虫脂肪含量最低,均为15%;幼虫期甘油含量随着龄期的增长而下降,从3.32μg/mg下降到1.12μg/mg,而蛹期逐渐上升,14日龄雌成虫甘油含量最高,为5.90μg/mg;1日龄幼虫总糖、海藻糖和糖原含量均最高,分别为7.51、0.94和1.93μg/mg,显著高于其他龄期瓜实蝇,蛹期糖类含量逐渐下降,而成虫期糖类含量随龄期增长而逐渐升高。【结论】本研究结果可为正确评估瓜实蝇的地理适应性和低温检疫处理提供依据。     

异色瓢虫幼虫对卵的种内自残和种间捕食

自残现象的发生在异色瓢虫Harmonia axyridis中很是常见,为明确自残行为对其存活和生长发育的影响,在非选择性条件下我们对异色瓢虫幼虫对非姊妹卵的种内自残和对七星瓢虫Coccinella septempunctata卵的种间捕食进行了研究。结果表明,异色瓢虫取食同种卵和异种卵均可完成生长发育,与取食蚜虫相比：①其从卵到成虫及蛹期的发育历期明显缩短（P<0.05）,其中1龄幼虫发育历期缩短非常明显（P<0.01）;②自然条件下高死亡率的1龄幼虫的存活率明显提高（P<0.05）;③2-4龄幼虫、蛹和成虫的体重变轻（P<0.05）,且体长在4龄幼虫间也表现出显著差异（P<0.05）。这表明异色瓢虫种内自残和种间捕食对其在食物恶劣条件下延续种群至关重要,但对种群的繁衍却不是最理想的途径。     

不同虫态及虫龄大草蛉与七星瓢虫之间的集团内互作

【目的】为明确新疆棉田棉蚜 Aphis gossypii 捕食性天敌之间的集团内捕食效应及其对蚜虫数量的控制作用。【方法】本研究以优势天敌昆虫大草蛉 Chrysopa pallens 和七星瓢虫 Coccinella septempunctata 为对象,以棉蚜为猎物,在温室中利用盆栽棉花,首先观察了2种天敌昆虫之间各虫态及虫龄配对的19个处理在无蚜植株上共存24 h后的存活数,然后观察了2种天敌昆虫配对处理下棉苗上棉蚜数量随时间的变化趋势。【结果】在无蚜棉株上2种捕食性天敌昆虫共存24 h后的存活结果表明：（1）在发育阶段相同的配对组合中,若是成虫则均存活,若是１龄幼虫则大草蛉存活较多,若是末龄幼虫则七星瓢虫存活较多;（2）在有卵的组配中,除七星瓢虫卵不被大草蛉成虫所捕食外,其他5个组配处理中卵均被捕食;（3）在有蛹的配对组合中,除七星瓢虫蛹被大草蛉末龄幼虫捕食外,其他处理下蛹均不被捕食;（4）在成虫与幼虫的配对组合中,七星瓢虫成虫捕食较多的大草蛉1龄幼虫,但不捕食大草蛉末龄幼虫,而大草蛉成虫与七星瓢虫1龄或末龄幼虫之间不发生捕食;(5）在不同龄期幼虫的配对组合中,大草蛉末龄幼虫捕食七星瓢虫1龄幼虫,而七星瓢虫末龄幼虫捕食大草蛉1龄幼虫。在有蚜植株上2种捕食性天敌共存对棉蚜数量具有不同的控制作用：（1）2种捕食昆虫的幼虫各自单独存在（对照）下,蚜虫密度随时间而降低;（2）大草蛉幼虫与七星瓢虫幼虫或成虫配对处理下,棉蚜密度随时间而增大;（3）大草蛉成虫与七星瓢虫幼虫或成虫配对处理下,棉蚜密度随时间而减小。【结论】研究结果说明,大草蛉与七星瓢虫之间存在集团内捕食,但２种天敌共存对棉蚜的控制作用取决于大草蛉虫态, 若大草蛉为幼虫,可使蚜虫密度增大,若为成虫,则使蚜虫密度减小。     

前裂长管茧蜂寄生行为对桔小实蝇幼虫生理指标的影响

通过观察测定了桔小实蝇幼虫生长发育过程中血淋巴蛋白种类和血细胞的变化以及前裂长管茧蜂的寄生行为对桔小实蝇幼虫各项生理指标的影响。结果表明:不同日龄桔小实蝇幼虫的血细胞浓度随着虫龄的增加呈显著上升趋势,由2龄的16.53×106cells/mL,到3龄的30.14×106cells/mL直至蛹前期的35.94×106cells/mL,但血淋巴蛋白种类没有明显变化。和未寄生幼虫相比,寄生后的幼虫各类型血细胞的浓度均下降,但差异均不显著;血淋巴蛋白种类无明显增减,但浓度有所变化;寄生4 h后血淋巴蛋白质浓度显著降低,接近22 h时升高,至化蛹前期浓度再次下降;寄生行为使幼虫的发育历期从8 d延长至9～11d;4日龄幼虫在被寄生后的第4 d起体重显著高于未被寄生的桔小实蝇幼虫。     

家蝇幼虫血细胞类型及免疫功能的初步研究

目的用不同方法观察家蝇3龄幼虫血细胞的形态,并对血细胞进行分类和免疫功能研究,为昆虫血细胞形态、分类及免疫研究提供实验依据。方法 (1)应用姬氏染色结合相差显微镜及荧光染色方法观察家蝇3龄幼虫血细胞形态,并对家蝇3龄幼虫血细胞进行分类;(2)观察家蝇3龄幼虫感染大肠杆菌后不同时间血细胞总数(THC)、各类血细胞数量(DHC)及形态的变化;(3)应用倒置显微镜观察家蝇3龄幼虫离体血细胞感染大肠杆菌后的形态变化;(4)采用酶细胞化学技术测定感染前后家蝇3龄幼虫血细胞中ACP、POD活性的变化。结果 (1)家蝇3龄幼虫血细胞可分为原血胞、浆血胞、粒血胞、珠血胞、类绛血胞5类,其中浆血胞又分为大核浆血胞和小核浆血胞两种;(2)感染后各时间组血细胞总数、浆血胞和粒血胞数量均显著升高,且浆血胞和粒血胞聚集成堆,出现细胞变形、空泡等形态变化;感染后16h、24h组的珠血胞数显著升高;原血胞和类绛血胞数量和形态无明显变化;(3)家蝇幼虫离体血细胞感染大肠杆菌后粒血胞周围见大量细菌聚集,浆血胞、粒血胞聚集成团将细菌包裹形成包囊,未见原血胞、珠血胞、类绛血胞形态的变化;(4)感染后浆血胞和粒血胞中ACP、POD的活性增强,感染前后原血胞、珠血胞、类绛血胞中均未见ACP、POD的阳性反应物。结论通过3种方法能很好地将家蝇3龄幼虫血细胞分为5类,其中浆血胞和粒血胞是家蝇幼虫参与免疫反应的主要细胞类型,珠血胞不参与感染后的早期细胞免疫反应。     

苍耳杆野螟生物学特性的研究

苍耳杆野螟是寄生在苍耳上的1种重要药用昆虫。在湖南（常德）每年发生4代,以老熟幼虫（五龄）在苍耳茎杆蛀道内越冬。室内自然变温下（22．2～25．0℃）饲养,卵期为4．8天,幼虫期为23．6天,蛹期为见11．8天,成虫期为6．1天。每雌可产卵124～585粒,块产。一、二龄幼虫食叶,三龄后蛀茎。     

A systematic study on hemocyte identification and plasma prophenoloxidase from Culex pipiens quinquefasciatus at different developmental stages

Culexpipiens quinquefasciatus (C. quinquefasciatus) is an important vector that can transmit human diseases such as West Nile virus, lymphatic filariasis, Japanese encephalitis and St. Louis encephalitis. However, very limited research concerning the humoral and cellular immune defenses of C. quinquefasciatus has been done. Here we present the research on hemocyte identification and plasma including hemocyte prophenoloxidase from C. quinquefasciatus at all developmental stages in order to obtain a complete picture of C. quinquefasciatus innate immunity. We identified hemocytes into four types: prohemocytes, oenocytoids, plasmatocytes and granulocytes. Prophenoloxidase (PPO) is an essential enzyme to induce melanization after encapsulation. PPO-positive hemocytes and plasma PPO were observed at all developmental stages. As for specific hemocyte types, prophenoloxidase was found in the plasmatocytes at larval stage alone and in the smallest prohemocytes during almost all developmental stages. Moreover, the granulocytes were PPO-positive from blood-fed female mosquitoes and oenocytoids were observed PPO-positive in pupae and in adult females after blood-feeding. As for plasma, there were different patterns of PPO in C. quinquefasciatus at different developmental stages. These results are forming a basis for further studies on the function of C. quinquefasciatus hemocytes and prophenoloxidase as well as their involvement in fighting against mosquito-borne pathogens.     

An immunological role of a PKC alpha binding protein (PICK1) expressed in the hemocytes of the beet armyworm, Spodoptera exigua

Protein kinase C (PKC) regulates various intracellular processes and its activity is tightly controlled by various factors, such as secondary messengers and binding proteins. A cDNA of a PKC alpha binding protein (also called PICK1: protein interacting with C kinase 1) was cloned in hemocytes of the beet armyworm, Spodoptera exigua (Noctuidae: Lepidoptera). It encodes 475 amino acid residues with putative PDZ and BAR domains interacting with other proteins or ligands. The PICK1 gene of S. exigua (Se-PICK1) was expressed in all developmental stages. In the larval stage, it was highly expressed in hemocyte and brain tissues. A quantitative RT-PCR indicated that its expression was significantly up-regulated by a bacterial challenge. RNA interference of Se-PICK1 in the fifth instar larvae with 100ng of a specific double-stranded RNA could effectively knockdown its expression after 48h post-injection in hemocytes. The suppressed expression of Se-PICK1 significantly impaired the larvae of S. exigua to induce hemocyte-spreading behavior and to form hemocyte nodules in response to bacterial infection. This is the first report of an immunological role of PICK1, which has been identified in various insect and mammalian genomes.     

苦瓜素Ⅰ对斜纹夜蛾几丁质酶基因(SlCht)和几丁质合成酶基因(SlCHS-A)表达及其生长发育的影响

【目的】几丁质酶和几丁质合成酶对昆虫的变态发育极其重要。本研究旨在阐明苦瓜素Ⅰ对斜纹夜蛾Spodoptera litura几丁质酶和几丁质合成酶基因表达及其生长发育的影响。【方法】利用RT-qPCR检测斜纹夜蛾几丁质酶基因(SlCht)和几丁质合成酶基因(SlCHS-A)在斜纹夜蛾不同发育阶段(卵、幼虫、预蛹、蛹和成虫)和4-6龄幼虫不同组织(体壁、中肠、脂肪体、血细胞、头部和马氏管)中的表达水平以及注射苦瓜素Ⅰ溶液(4 μg/头) 24, 48和72 h时斜纹夜蛾SlCht和SlCHS-A在6龄幼虫各组织中的表达水平。分析在斜纹夜蛾4龄幼虫中注射不同浓度(31.25, 62.5, 125, 250和500 ng/头)的苦瓜素Ⅰ溶液对幼虫和蛹历期、幼虫增重、蛹重、蛹长度、化蛹率、羽化率和存活率的影响,并利用体视显微镜观察斜纹夜蛾幼虫的表型变化。【结果】SlCht和SlCHS-A在斜纹夜蛾中的表达具有发育阶段特异性。SlCht和SlCHS-A在卵期表达量最高,幼虫期和预蛹期的表达量较低;在各幼虫期又表现为6龄幼虫期表达量最高,在其他龄期的表达量低。SlCht和SlCHS-A在斜纹夜蛾6龄幼虫中也显示出组织特异性表达,在血细胞和体壁中高表达,在头部、中肠和脂肪体中低表达。在斜纹夜蛾6龄幼虫中注射苦瓜素Ⅰ能诱导SlCht和SlCHS-A在其各组织中表达量降低;在4龄幼虫中注射苦瓜素Ⅰ后,斜纹夜蛾的生长发育受到抑制,幼虫增重延缓,发育历期延长,化蛹率下降甚至化蛹失败,幼虫及蛹出现较高的畸形率。【结论】苦瓜素Ⅰ可通过诱导SlCht和SlCHS-A表达量的降低来实现对斜纹夜蛾生长发育的抑制作用。本研究为进一步阐明苦瓜素对斜纹夜蛾生长发育的抑制机制提供了新的理论基础,并为进一步应用苦瓜素Ⅰ进行防控奠定了基础。     

Hemocytes of the blowfly Calliphora vicina and their dynamics during larval development and metamorphosis

On the basis of in vitro observation of live cells and examination of stained slides of larval and prepupal Calliphora vicina hemolymph, seven types of hemocytes have been detected: prohemocytes, stable and unstable hyaline cells, thrombocytoids, spindle cells, larval plasmatocytes, and plasmatocytes I-IV, a. The last representing sequential stages of one cell line differentiation. Prohemocytes are basic cells, from which other forms of hemocytes derive outside the hemopoietic tissue, i.e. in free hemolymph. At the last larval instar, three waves of hemopoiesis occur. Either wave tends to increase the general number of cells and to change the quality of hemocyte population. The first wave occurs at the close of larva feeding and is accompanied by increase in the number of hyaline hemocytes, thrombocytoids and larval plasmatocytes. The second wave of hemopoiesis occurs after the larva's crop emptying. In this period the main increase of hemocyte population occurs at the expense of prohemocytes and plasmatocytes I. The most significant (five-fold) explosion of the population of free hemocytes takes place at the onset of pupariation and correlates with the rise of ecdysone titer. At the first stage of this peak, the amount of plasmatocytes I sharply increases. Further on these are rapidly differentiated into plasmatocytes II and III. After the puparium formation, hemocytes are reduced in number. Plasmatocytes III phagocytose fragments of destroyed larval tissues, pass to the stage of plasmatocytes IV (macrophages), and partially settle on tissues.     

Ultrastructure of the hemocytes of Cetonischema aeruginosa larvae (Coleoptera,Scarabaeidae): involvement of both granulocytes and oenocytoids in in vivo phagocytosis

In the context of comparative studies on immunity defence mechanisms of adults and larvae of the coleopteran Cetonischema aeruginosa (Drury, 1770) the ultrastructure of the circulating hemocytes of the third instar larval stage has been investigated by means of light and transmission electron microscopy (TEM). Six types of hemocytes were found in the hemolymph of C. aeruginosa and they were identified as prohemocytes, granulocytes, plasmatocytes, coagulocytes, oenocytoids and spherule cells. In order to identify the "professional" phagocyte cell, phagocytosis assays were performed in vivo by injection of 0.9 microm carboxylate-modified polystyrene latex beads. It was demonstrated that the granulocytes and the oenocytoids of C. aeruginosa were the only hemocyte types involved in this cellular response.     

Antibacterial activity of silkworm hemolymph after hemopoietic organ being injured with radiosurgery

Abstract:  To study the effect of hemopoietic organs damage on hemocyte function and antibacterial activity of hemolymph, silkworm ( Bombyx mori ) larvae were locally irradiated with carbon ion beams (¹²C⁵⁺, 100 Gy), live and death ratio of hemocytes and antibacterial activity of hemolymph were investigated. For unirradiated controls, the ratio of died hemocytes hardly changed at the fifth instar, but for locally irradiated silkworms, with growth died hemocytes and low-functional hemocytes increased clearly, and reached an extremely significant level at the later stage of the fifth instar. For irradiated individuals, the phenolxidase activities and sterilization effect of hemolymph were clearly lower than those of controls. So it is considered that after irradiating hemopoietic organs with heavy ion beams, not only the number of hemocytes decreased but the function of hemocytes also dropped, and they at last lead to a decline in immunity.     

Hematopoiesis in larval Pseudoplusia includens and Spodoptera frugiperda

Maintenance of circulating hemocytes in larval Lepidoptera has been attributed to both mitosis of hemocytes already in circulation and the release of hemocytes from hematopoietic organs. In this study, we compared hematopoiesis in the noctuids Pseudoplusia includens and Spodoptera frugiperda. For both species, hemocyte densities per microl of blood increased with instar. Differential hemocyte counts indicated that plasmatocytes were the most abundant hemocyte type during early instars but granular cells were the most abundant hemocyte type in the last instar. Hematopoietic organs were located in the meso- and metathorax of S. Frugiperda and P. Includens. These organs contained large numbers of hemocytes in S. Frugiperda, but contained few hemocytes in P. Includens. The majority of the hemocytes recovered from hematopoietic organs were identified as plasmatocytes. Using hemocyte type-specific markers and bromodeoxyuridine (BrdU) incorporation experiments, we determined that all hemocyte types with the exception of oenocytoids synthesize DNA. BrdU labeling indices for both species also fluctuated with the molting cycle. Ligation experiments suggested that hematopoietic organs are an important source of circulating plasmatocytes in S. Frugiperda but not in P. Includens. Injection of heat killed bacteria into larvae induced higher levels of BrdU labeling than injection of sterile saline, suggesting that infection and wounding induce different levels of hemocyte proliferation. Arch.