长江下游地区4个克氏原螯虾群体的遗传多样性分析

为了对长江下游地区克氏原螯虾(Procambarus clarkii)群体的遗传多样性状况进行本底调查, 本文利用17对微卫星引物对南京(NJ)、盱眙(XY)、合肥(HF)、南昌(NC)4个克氏原螯虾地理群体进行了遗传多样性研究。结果表明, 4个地理群体的平均观测杂合度为0.4322–0.4826、平均期望杂合度为0.4024–0.6121、平均多态信息含量为0.3408–0.5624, 4个克氏原螯虾群体的遗传多样性处于中等水平, 其中南京群体遗传多样性最高, 合肥群体最低; 4个克氏原螯虾群体的基因流为1.3729–5.9161, 遗传分化指数为0.0405–0.1540, 群体间基因流水平较高, 群体间遗传分化程度较小。聚类结果显示4个群体可分为2支: 南京、南昌、盱眙三个群体聚为一支, 合肥群体独自成一支。以上结果将为克氏原螯虾遗传育种、资源保护和利用提供一定的理论依据。     

桂林地区克氏原螯虾对泽蛙蝌蚪的捕食

克氏原螯虾（Procambarus clarkii）已入侵我国江苏、湖北、江西、安徽等多个省（市）。为研究克氏原螯虾对其生境中主要两栖动物泽蛙（Rana limnocharis）的影响,我们于2006年5-6月对广西桂林地区自然生境中克氏原螯虾和泽蛙蝌蚪的数量和密度进行了调查,7月在室内进行了克氏原螯虾对泽蛙蝌蚪和饰纹姬蛙（Microhyla ornata）蝌蚪的捕食实验。野外调查结果表明克氏原螯虾的密度与泽蛙蝌蚪的密度呈显著负相关。室内实验结果表明克氏原螯虾对泽蛙蝌蚪的捕食强度与克氏原螯虾体长呈显著正相关,且对泽蛙蝌蚪的捕食强度高于饰纹姬蛙蝌蚪。表明克氏原螯虾对两栖类幼体有比较严重的危害,应加强监测,采取相应措施预防和控制其危害。     

淮河克氏原螯虾种群遗传结构

克氏原螯虾（Procambarus clarkii）是广泛分布于我国淡水水域的外来入侵物种,对其种群遗传结构的研究有助于了解种群扩散过程。本文测定了淮河流域自然水体中9个地理种群共151个野生克氏原螯虾线粒体COⅠ序列,获得624 bp的有效基因片段,共定义了25个单倍型。AMOVA分析结果表明,淮河流域的克氏原螯虾种群内的变异占主导地位,单倍型多样性和核苷酸多样性均为中等水平,各地理种群间平均遗传差异较小（F_st=0.078）,遗传差异不显著。系统发生树并没有按地理位置形成对应族群,表明克氏原螯虾各地理种群间存在明显的基因流,各地理种群间没有形成明显的种群遗传分化。     

克氏原螯虾的入侵生态学研究进展

克氏原螯虾(Procambarus clarkii)原产于美国南部和墨西哥北部,是一个著名入侵物种,作为一种水产经济资源物种在世界各地扩散。克氏原螯虾抗逆性强,所具有的广泛生境适应性、生长迅速、高生殖率等特点使它们迅速建立野生种群。近10余年的研究认为,克氏原螯虾通过捕食和资源竞争等机制严重威胁引入地的水生植物、无脊椎动物、两栖类等的生存,显著降低引入地的生物多样性。当前,由于克氏原螯虾的经济价值高,它会借助于人力的作用而继续扩散。为认清和减少克氏原螯虾对引入地的生态影响,应加强以下方面的研究:1)在中国开展克氏原螯虾的生态危害的调查和研究;2)克氏原螯虾种群调节和控制对策研究;3)被入侵地的生态恢复工作。     

外来种克氏原螯虾的危害及其防治

克氏原螯虾作为外来入侵种,已经对其所入侵的栖息地土著动物、植被、土壤结构等生态环境构成了严重威胁。针对我国一些拟引入养殖或正在大面积推广养殖该螯虾的现状,提出防治该物种的一些探索性对策,对拟将引种的地区,必须持慎重态度,进行生态安全评估,制定预警措施     

安徽地区克氏原螯虾白斑综合征的诊断及朔源

[目的]诊断安徽地区养殖的克氏原螯虾发生疫情的病原,追踪感染源,为疫病的防控提供依据.[方法]采集病料,参照OIE推荐的套式PCR法检测白斑综合征病毒(WSSV).将病螯虾鳃丝组织研磨后离心,取上清液接种健康克氏原螯虾.透射电镜观察病原特征.综合流行病源学及病原检测,分析养殖地区的可能感染源.[结果]发病螯虾感染了WSSV,动物实验可见和发病螯虾相同的临床症状,证实该病原引起养殖螯虾发病.电镜观察可见典型的白斑综合症病毒颗粒.流行病学调查显示,水产颗粒膨化饲料和冷冻饲料甲壳类检测阳性.[结论]该地区养殖的克氏原螯虾近年爆发的疫病病原为WSSV,推断地区性克氏原螯虾WSSV的感染并流行与污染的饲料有关.     

武汉地区克氏原螯虾繁殖期的研究

为探究武汉地区克氏原螯虾(Procambarus clarkii)繁殖期的问题,采用石蜡切片的方法,对武汉地区自然群体中克氏原螯虾雌雄成体的生殖腺进行了周年观察研究。结果表明,武汉地区成年克氏原螯虾的卵母细胞发育不同步,在4~5月以及9~10月的卵巢中都有大量成熟期的卵子,同时也含有少量卵黄发生前期或初期的卵母细胞。6~7月和11月都可见产后恢复期的卵巢,内含少量未产出的正在退化的过熟卵和一些卵黄发生前期或初期的卵母细胞。精巢发育的年周期变化没有卵巢变化的那么明显,但也可以看到,在4~6月和9~10月,含精子细胞和精子的精小管的比例大大增加,而在其他时期,含精原细胞和精母细胞的精小管比例较多。这些结果提示,武汉地区克氏原螯虾一年有两个繁殖高峰期,一个在4~5月,另一个在9~10月。     

圈养小熊猫遗传多样性与种群遗传结构分析

小熊猫是亚洲特有的珍稀濒危动物,目前受到栖息地减少、片断化和人类活动干扰等威胁。中国圈养小熊猫已经有60 多年历史,约55 个机构曾经饲养过小熊猫,现今圈养数量有400 多只,评估小熊猫圈养种群的遗传多样性和遗传结构对科学维持圈养种群和保存遗传种质资源意义重大。本研究利用19 个微卫星座位,对中国境内11 个小熊猫圈养种群的116 只个体进行了遗传多样性评估及遗传结构分析。结果显示11 个种群都具有较高的遗传多样性,平均基因丰富度3.505 ± 1.033 (北京)至4.026 ± 1.219 (冕宁),期望杂合度0.631 ± 0.225(黄山)至0.782 ±0.171 (温岭)。其中福州和无锡种群极显著偏离Hardy-Weinberg 平衡。整个圈养群体内各个种群遗传分化系数为0.055,呈显著分化,表明11 个种群遗传分化水平较高。Bayesian 遗传聚类分析将11 个种群聚为三个遗传簇,与野生种群的遗传聚类结果一致。结论:小熊猫圈养种群与野生种群相比,同样具有较高的遗传多样性。因此,圈养小熊猫遗传管理的重点不再是引进野生个体充实圈养种群,应制订科学的繁殖计划,避免近交,从而维持圈养种群的遗传多样性。     

仙姑弹琴蛙蝌蚪对陌生捕食者克氏原螯虾的反捕反应

以峨眉山的仙姑弹琴蛙 (Ranadaunchina)蝌蚪作为猎物 ,该地区潜在的入侵物种克氏原螯虾 (Procambarusclarkii)为捕食者 ,于冬季设计实验观察仙姑弹琴蛙蝌蚪对陌生入侵捕食者的反捕行为反应。实验按照 3× 3× 2因子设计 ,即 :捕食者状况 3个水平 (食植物的克氏原螯虾、食蝌蚪的克氏原螯虾和无捕食者 ) ,蝌蚪年龄组 3个水平(31- 35期的 2 .5龄蝌蚪、2 6 - 30期的 1.5龄蝌蚪和 2 5期的 0 .5龄蝌蚪 ) ,以及蝌蚪短期经验 2个水平 (有经验和无经验 )。实验发现 :有短期被捕食经验的蝌蚪的活动时间极显著低于无被捕食经验的蝌蚪 ,而静止时间以及隐蔽所利用时间却极显著地高于无经验的蝌蚪 ;短期经验和捕食者状况的交互效应对蝌蚪的活动时间有显著影响 ;而蝌蚪年龄、捕食者状况及其交互效应对蝌蚪的行为反应没有显著影响。这表明 ,有短期被捕食经历的仙姑弹琴蛙蝌蚪并未通过个体学习建立起对陌生捕食者克氏原螯虾的识别能力 ,而是表现出无针对性的活动性受抑、隐蔽时间增加的行为反应。如果自然条件下的仙姑弹琴蛙蝌蚪也有类似反应 ,在克氏原螯虾入侵初期 ,仙姑弹琴蛙蝌蚪则可能会由于无法识别新的捕食者而表现出过度的反捕行为反应 ,承受更大的亚致死作用 ,而对种群生存造成不利影响     

蒙新区子午沙鼠种群的遗传多样性和遗传结构

为了探讨蒙新干旱区景观和环境因素对子午沙鼠种群遗传结构的影响,我们利用8 个多态性的微卫星位点,对子午沙鼠14 个局域种群的160 个个体进行了种群遗传多样性和遗传结构的分析。微卫星结果显示种群具有高的遗传多样性,微卫星位点的等位基因数(A)为22. 50 ±3.02 (19 ～ 28),多态信息含量(PIC)为0.912 ±0. 02 (0.872 ～ 0. 929);种群观测杂合度(HO )为0.68 ± 0. 19 (0. 52 ～0.85),期望杂合度(HE )为0.79 ± 0. 08(0. 71 ～ 0.85)。分子变异分析(AMOVA)结果显示,所得到的3 个组之间已发生显著的遗传分化(P < 0. 001)。
Mantel 检测结果表明,子午沙鼠种群遗传结构与地理距离无显著的相关性(P = 0.270 0),而在多元线性模型中,蒙新干旱区的海拔高度是影响种群遗传结构的关键因素。     

克氏原螯虾(Procambarus clarkii)血细胞染色方法的研究

甲壳类血细胞的形态和分类是甲壳类免疫学研究的基础.本文选择三种甲壳类血细胞的常用染料:瑞氏染液、姬姆萨染液和瑞氏-姬姆萨混合染液,研究它们对于克氏原螯虾血细胞的染色效果.通过改变染色时间、染色温度、分色方式以及分色时间等染色条件,观察、比较不同染色条件下血细胞内的颗粒、细胞核和细胞质的着色情况,以及细胞整体轮廓清晰程度,确定适用于克氏原螯虾血细胞染色的理想染色方法,并建立相应的操作程序.     

Invasion of the shelter snatchers: behavioural plasticity in invasive red swamp crayfish,Procambarus clarkii

1. In many freshwater systems, competition for shelter plays an important role in determining the persistence of both native and alien species. The red swamp crayfish, Procambarus clarkii, is currently invading the native habitat of the signal crayfish, Pacifastacus leniusculus, in southern Oregon, and interspecific competition for shelter may be driving the species replacement in this region. 2. We designed a 2 × 3 factorial mesocosm experiment, with shelter density and species combination as factors, to investigate shelter occupancy and resource competition. Contrary to our predictions, the two crayfish species are equal competitors for shelter. Further, the invasive P. clarkii modified its shelter occupancy behaviour in the presence of the native P. leniusculus and has broader microhabitat preferences. 3. Specifically, we found that P. clarkii alters shelter occupancy and space use patterns when the two species occurred together, such that shelter use was identical between P. clarkii and P. leniusculus in mixed‐species treatments. In such treatments, both species increased their use of shelters when shelter density increased. When P. clarkii was alone, however, individuals did not alter shelter use as a function of shelter density, whereas P. leniusculus exhibited similar density‐dependent behaviour in both mixed‐ and single‐species treatments. 4. In a complementary field survey, we employed an ‘epicentre‐based’ design to sample two field sites. We observed patterns of microhabitat use and breadth for each species similar to those in our mesocosm experiment: the invasive P. clarkii was more abundant across different habitats and used a broader range of microhabitats than the native P. leniusculus. As such, we found that P. clarkii was more abundant across both field sites than the P. leniusculus, occupying microhabitats within and beyond the preferred range of P. leniusculus. Both field sites were affected by urban development and agriculture. 5. The use of microhabitats by both species was similar in the laboratory and the field. This study confirms that P. clarkii individuals can, and do, successfully occupy microhabitats preferred by P. leniusculus in the Willamette Valley. The results from our study may be relevant to other freshwater systems inhabited by P. clarkii and contribute to the understanding of ‘niche opportunity’, a concept which defines the environmental conditions that promote biological invasions.     

Crayfish invading Europe: the case study of Procambarus clarkii

The red swamp crayfish, Procambarus clarkii, native to northeastern Mexico and southcentral USA, is today the dominant macroinvertebrate in several European countries. While the first introduction of this species into Spain is well-documented, little is known about its pathways of invasion and the reason for its rapid spread in several European countries. Study of the biology of the species has revealed a number of properties that makes this crayfish a successful invader. Procambarus clarkii exhibits properties characteristic of an r-selected species, including early maturity at small body size, rapid growth rates, large numbers of offspring at a given parental size, and relatively short life spans. It is also plastic in its life cycle, able to disperse widely in the habitat and to tolerate environmental extremes. It displays generalist and opportunistic feeding habits, consuming macrophytes and preying on amphibians. Procambarus clarkii can also replace indigenous crayfish by a combination of mechanisms, including competitive exclusion and transmission of the fungus-like Aphanomyces astaci, responsible for the crayfish plague. Finally, this species shows a wide behavioral flexibility when coping with new types of predators. The results of these studies, combined with the increasing information available in the scientific literature on this and other crayfish species, will help us understand invasions in this taxon and make predictions about the identity of future crayfish invaders.     

Habitat use and dispersal of the invasive crayfish Procambarus clarkii in ephemeral water bodies of Portugal

We used radio-telemetry to analyze habitat use and dispersal of the invasive crayfish Procambarus clarkii in the extreme environmental conditions of a temporary river in southern Portugal. The rationale of this study was that an understanding of the properties that make this species a successful invader can be of help for the conservation of freshwater biodiversity. The results showed that: (1) crayfish displaced at a similar rate (ranging 2.5-38 m d^-1) as other European populations; (2) movement was faster at night; (3) crayfish movement had no clear environmental correlates, except for water temperature; (4) peaks of more intense locomotion were intercalated with longer periods of slow or null speed; and (5) P. clarkii did not excavate burrows in the study habitat, but took refuge under boulders and most often occupied complex microhabitats (e.g., vegetated sections of the river). The conclusion was that, notwithstanding their extreme conditions, ephemeral water bodies in southern Europe are highly susceptible to invasion by P. clarkii.     

克氏螯虾虾壳制备氨基葡萄糖盐酸盐的工艺研究

研究了以克氏螯虾壳为原料制备氨基葡萄糖盐酸盐 (GAH)的工艺条件 :酸水解液为 10mol·L-1HCl,85～ 92℃水解 5～ 6h ;甲壳素∶酸水解液为 3∶5。GAH产率为甲壳素的 5 2 %～ 5 5 %。GAH纯度 :99%～ 10 0 .5 %。     

克氏原螯虾精巢与输精管组织细胞原代培养方法初探

将剪碎的克氏原螯虾(Procambarus clarkii)精巢与输精管组织块分别接种于含20?S、双抗以及添加了适量无机盐等的M199-MK、MEM、S20、Grace’s和L-15培养基中,25℃恒温静置培养。结果表明,在Grace’s培养基中,只有少量组织块贴壁。在其余培养基中,大多数组织块能在1～2周内逐渐贴壁,L-15和S20培养基培养效果最好。组织块贴壁后,一些成纤维细胞和上皮样细胞逐渐从组织块迁移出来。传代后组织块可继续贴壁并重新迁移出一些成纤维细胞与少量上皮样细胞。添加部分草鱼细胞系PSF或斜纹夜蛾细胞系Sl驯化的培养基进行培养,驯化培养基:新鲜培养基=1:5(v/v),结果表明,Sl驯化培养基能促进细胞的迁移。组织块一般可离体培养2～4个月,但是都未能形成细胞单层。胰蛋白酶消化后分散的虾细胞不能贴壁。     

Methyl farnesoate controls adult male morphogenesis in the crayfish, Procambarus clarkii

Adult male crayfish Procambarus clarkii exist in two morphotypes. They continue to molt as adults, switching between Form Is and Form IIs. Form Is are primary reproductive types, with large chelae and spines on the ischiopodites of the third and fourth pair of walking legs. Form IIs are non-reproductive types with smaller chelae and no spines on the ischiopodites. We investigated the hormonal control of these transitions in two ways, by eyestalk ablation and by methyl farnesoate (MF) treatments. Eyestalk ablation accelerates molting and increases MF levels in the blood. MF is a hormone that regulates both reproduction and morphogenesis. MF concentrations were determined in two ways. The hemolymph samples were extracted first, then purified, using normal phase HPLC. The fractions containing MF were collected and analyzed for MF concentration, utilizing both internal and external standards by GC/MS. The other hemolymph samples were analyzed from individual animals by HPLC. The concentrations of ecdysteroids were determined by radioimmunoassay. In the control animals, 4 out of 4 untreated Form I males molted into Form II, while 6 out of 7 Form IIs molted into Form Is. Eight of 8 ablated Form Is molted into Form IIs as expected, while 5 of 5 ablated Form IIs molted into Form IIs, instead of Form Is. MF treatment of intact animals resulted in 6 of 7 Form Is becoming Form IIs and 5 of 6 Form IIs becoming Form IIs. These results were highly significant in comparison of Form I and IIs in each treatment (eyestalk intact, eyestalk ablated and eyestalk intact with MF) by a chi square analysis, P = 0.006, P < 0.0005, and P = 0.013, respectively. MF premolt blood levels suggested that Form IIs were produced in the presence of 1.3 ng/ml MF, while Form Is result from MF levels less than 0.5 ng/ml. Since both eyestalk ablation and MF treatment resulted in the failure of Form IIs becoming Form Is, it was concluded that the control of morphogenesis of primary reproductives (Form Is) depends on a low level of MF prior to the molt, while Form IIs are formed in the presence of increased levels of MF.     

切除眼柄对加速克氏原螯虾卵巢发育进程的组织学研究

应用组织切片和透射电镜技术研究眼柄切除对加速克氏原螯虾(Procambarus clarkii)卵巢发育进程的影响。实验分成4组,选取眼柄切除15 d和30 d两个时间点进行。切除眼柄30 d后,雌虾体长增长显著,从(11.50±2.30)cm增长至(18.20±3.40)cm(P0.05),性腺指数(GSI)从4.56%±2.81%增长至8.05%±2.51%(P0.05),体重和螯肢长也有所增长(P0.05)。电镜观察发现卵母细胞从发育Ⅱ期迅速度过Ⅲ期进入到Ⅵ期阶段,卵母细胞体积变大,变圆;液泡压缩变小;卵黄颗粒加速积累,逐渐占满整个胞内空间;滤泡细胞胞质从黏稠变稀薄。这些结果表明,切除眼柄在短期内显著加快卵母细胞内卵黄颗粒的积累,并且加快个体生长。