Estudo in vitro de derivados sintéticos da Isoniazida e da Pirazinamida sobre Leishmania (Viannia) braziliensis

As leishmanioses são um grupo de doenças causadas por protozoários do gênero Leishmania spp que afetam 98 países. No Brasil, no ano de 2013, foram relatados 3.253 casos de leishmaniose visceral e 18.226 casos de Leishmaniose Tegumentar Americana. O tratamento de primeira escolha continua sendo realizado com antimoniais pentavalentes, e em casos de insucessos os fármacos de segunda escolha são a pentamidina e a anfotericina B. Tais medicamentos causam intensos efeitos adversos e ultimamente têm surgido cepas resistentes aos mesmos. Em áreas endêmicas têm sido cada vez mais comum o surgimento da co-infecção Leishmania com Mycobacterium tuberculosis. O tratamento para a tuberculose com pirazinamida (PZA) e isoniazida (INZ), controla a leishmaniose. Esses dados sugerem atividade anti-leishmania da PZA e da INZ. O objetivo deste trabalho foi avaliar a atividade in vitro da INZ e da PZA e seus compostos derivados (série G e série R, respectivamente) sobre Leishmania (Viannia) braziliensis. As moléculas foram testadas em monocamadas de macrófagos peritoneais de camunongos infectados com L. (V) braziliensis durante 48h. Todas as moléculas testadas inibiram o índice de infecção de forma dose dependente em comparação aos controles. As moléculas da série R foram mais ativas do que a PZA, porém o resultado foi significativo somente para a R02 (p < 0,005). Apenas a molécula R05 (76,64M) foi relativamente tóxica para macrófagos. Os compostos mais ativos foram R02, G01 e G02, cujos índices de seletividade foram 14,31, 19 e 30, respectivamente. A dosagem de nitrito foi feita em sobrenadantes de monocamadas de macrófagos peritoniais infectados e tratados com as substâncias nas concentrações 10 e 100M. A G01 e a G02 estimularam a produção de NO2 nas duas concentrações, entretanto o resultado foi estatisticamente significativo para a G02 em 100M (p < 0,0001), a G05 só estimulou óxido nítrico na maior concentração. Todos os compostos da série R estimularam NO2, contudo, o resultado foi estatisticamente significativo para a R03 e R05 a 100M (p < 0,001). Adicionalmente, foi realizado uma análise preditiva in sílico de parâmetros farmacocinéticos das moléculas mais ativas in vitro, utilizando o software admetSAR. Os dados obtidos mostraram que de forma semelhante às suas moléculas originais a G01, G02 e R02 apresentaram alta capacidade de serem absorvidas pelo trato gastrointestinal, baixo potencial hepatotoxico e carcinogênico. Juntos, esses dados demonstram que essas moléculas são seletivamente tóxicas para o parasito com potencial para serem testadas pela via oral em estudos em modelo experimental de infecção.

Análise comparativa do reconhecimento celular de antígenos dos subgêneros de Leishmania na leishmaniose tegumentar americana

Células Mononucleares do sangue periférico (CMSP) oriundas de 28 pacientes que residem em áreas endêmicas para Leishmania braziliensis foram estudados. Destes, 10 encontravam-se na fase ativa da doença e 18 curados clinicamente para as lesões de leishmaniose. As células foram cultivadas frente à antígenos de 6 espécies de Leishmania: Leishmania: L. (V.) braziliensis (LbAg), L. (V.) guyanensis (LgAg), L. (V.) lainsoni (LlAg), L. (V.) naiffi (LnAg), L. (L.) amazonensis (LaAg), L. (L.) chagasi (LcAg) e antígeno de concanavalina A como mitógeno. Os resultados foram expressos em índices de estimulação (IE). Temendo uma possível degradação protéica, os antígenos foram preparados com PBS e com PBS tampão antiproteolítico e o perfil eletroforético foi analisado através de SDS-PAGE. Os sobrenadantes das culturas foram estocados para os ensaios de ELISA para detecção de IFN-γ. A análise do perfil eletroforético dos antígenos de diferentes espécies de Leishmania demonstrou um padrão distinto e heterogêneo e aparentemente, não foi observada degradação assim que os antígenos foram preparados e 10 meses de estoque. Entretanto bandas parecem ser compartilhadas entre as espécies, que podem estar associadas a uma conservação de antígenos entre as espécies de Leishmania. A resposta proliferativa dos linfócitos (RPL) dos pacientes da forma ativa foi mais intensa para LbAg (1710,3), seguido de LnAg (177,6), LaAg (16,89,8) e LlAg (1410,1), e menos intenso para LgAg (11,46,6). Quando os IE obtidos para LbAg foram comparadas com outras espécies observou-se diferenças para LgAg (p<0.004) e LlAg (p<0.03). Não foram encontrados diferenças no perfil protéico dos extratos antigênicos produzidos com e sem inibidores de protease. Os estímulos obtidos de CMSP de indivíduos curados clinicamente não demonstraram diferenças quando comparados com aqueles da fase ativa da doença. O índice de estimulação (médiadesvio padrão) frente estímulos antigênicos pouco variou entre as espécies: LbAg - 22,222,2, LnAg - 15,911,5, LgAg - 20,720,6, LlAg - 14,710,1, LaAg - 15,1114,5 e L. chagasi - 13,78). A produção de IFN-γ quantificada nos sobrenadantes das culturas frente aos antígenos totais e solúvel mostrou níveis da citocina mais elevados quando utilizou-se antígenos total de L. guyanensis (568,1544,5; mediana: 518,5 pg/mL) quando comparadas às outras espécies analisadas. Porém quando analisadas o antígeno solúvel, podemos observar que tanto para os antígenos da espécie L. braziliensis quanto para L. naiffi, a fração total obteve os maiores índices de estímulos quando comparadas com a fração solúvel. Estes estudos sugerem que há uma reatividade cruzada entre as espécies dos subgêneros Viannia e Leishmania.

苔藓植物一些东亚特有属的分子系统学研究

苔藓是高等植物（有胚植物或陆地植物）中最原始的一类，但种类却丰富多样，其形态和生长环境的多样化程度高于蕨类和裸子植物，且对极端环境的忍耐力更强，分布范围也更广。“特有”是一个地理概念，它是相对广布而言，当一个类群的分布范围有一定的限制时即为特有现象。“东亚特有”是指分布范围主要局限于中国，朝鲜，日本和蒙古等，向北可及俄罗斯远东地区，少数可分布至中国南部相邻地区的植物类群。东亚地区主要以温带植物区系为主，但也包含一些热带植物区系成分，还因为第四纪以来受冰川活动影响较少，因此植物种类非常丰富。东亚地区也是苔藓植物的多样性中心之一，这里有较多的特有成分。在我国总共分布有苔藓植物东亚特有属35属，其中苔类5属，藓类30属。长期以来，特有成分始终引起人们的极大关注，不仅是因为其在植物地理学上的重要性，还因为特有类群中包含了孓遗类群，往往系统位置比较关键，此外，大部分特有类群对人为干扰比较敏感，对其保护就愈加重要，因为它在这个地区的消失就意味着一个类群的灭绝。 我国对苔藓植物东亚特有类群已有较好的认识，在前人知识积累的基础之上，我们期望通过分子系统学的方法，开展对东亚特有苔藓属的研究，逐步揭开特有属植物的神秘面纱，最终在系统树上找到它们各自应该属于自己的位置。 在本次研究中，我们总共得到十一个苔藓植物东亚特有属的新鲜材料。在实验室中我们对这十一个特有属叶绿体和核的六个基因（叶绿体atpB, rbcL, cp-SSU, cp-LSU 和核18S，26S rDNA）进行了测序，并在此基础之上，构建了来自苔藓植物106个属上述六个基因的联合矩阵，并对它们进行了系统学分析。本文所选十一个特有属中除三个苔类属和一个线齿藓类的属之外，其它七个特有属都属于侧蒴藓类。根据近几年的研究结果，侧蒴藓类中灰藓目被认为是起源自一次快速辐射演化，灰藓目各科之间的关系以及各科的范围都很难确定。即便本实验测序一万多bp，这一支之内的关系仍不能解决。 在以上结果的基础上，本文对线齿藓类的树发藓属（Microdendron）进行了较为详细的研究，我们用最大简约法分析了金发藓目15属，33种的18S, rbcL和trnL-F序列的联合矩阵。对树发藓属的微形态进行了电镜扫描。形态和分子数据的分析结果表明，这个特有属在属级水平是不成立的，它仅是小金发藓属的一个种。此结果支持将这个东亚特有属降为种的等级。此外，本文还对囊绒苔属（Trichocoleopsis）和新绒苔属（Neotrichocolea）的系统位置做了比较详细的研究。我们分别分析了一个苔类植物57属的四基因（cp-SSU, cp-LSU, atpB and rbcL）矩阵和一个苔类植物24属的九基因（cp-SSU, cp-LSU, atpB, psbA, rps4, rbcL, 18S, 26S and nad5）联合矩阵，结果显示囊绒苔属和新绒苔属互为姐妹群关系，而毛叶苔属（Ptilidium）又是它们二者的姐妹群。研究结果支持了囊绒苔属和新绒苔属组成新绒苔科（Neotrichocoleaceae），而不同于前人的观点：将上述两属放置于毛叶苔科（Ptilidiaceae）、绒苔科（Trichocoleaceae）或多囊苔科（Lepidolaenaceae）。另外值得注意的是这两个特有属和毛叶苔属组成的一支位于叶苔类（Leafy liverwort）中“Leafy I”和“Leafy II”两大支之间，但这一支确切的系统位置没有解决，仍有待于进一步研究。 除此之外，本文还利用GenBank中的数据对东亚特有属日鳞苔属（Nipponolejeunea）和耳坠苔属（Ascidiota）（未获得实验材料）进行了初步的系统学分析。结果表明传统上放在细鳞苔科的日鳞苔属与毛耳苔科的毛耳苔属（Jubula）为姐妹群关系，建议将日鳞苔属置于毛耳苔科;耳坠苔属是光萼苔科的成员，属的分类等级是合理的。 最后本文利用罚分似然法，选取多个化石作为标定点，对来自苔藓植物主要类群及其它陆地植物共115个类群5个基因（atpB, rbcL, cp-SSU, cp-LSU, 18S）的矩阵进行了分子钟的分析，初步估算11个东亚特有属的分化时间。

鳞毛蕨科、叉蕨科和藤蕨科的分子系统学研究

利用叶绿体rbcL和atpB基因对鳞毛蕨科、叉蕨科和藤蕨科进行了系统发育重建。对鳞毛蕨科的范畴和科下划分进行了重点研究，同时基于叶绿体rps4-trnS和trnL-F序列对该科三个东亚特有属（玉龙蕨属、柳叶蕨属和鞭叶蕨属）的分类等级进行了研究，基于rbcL、atpB和accD基因重新讨论了拟贯众属的系统位置。主要内容包括： 1.鳞毛蕨科的界定及三个东亚特有属的系统位置 利用rbcL和atpB基因，探讨了鳞毛蕨类及其近缘类群的系统发育关系。取样包括了鳞毛蕨类所有主要的分类群，尤其是增加了中国和东亚地区的类群代表。两个基因片段的联合分析较好地解决了鳞毛蕨类及其近缘类群的系统发育关系。研究结果显示广义鳞毛蕨科是个多系类群，传统上置于鳞毛蕨科中的蹄盖蕨类athyrioid、球子蕨类onocleoid和叉蕨类植物tectarioid均应该从鳞毛蕨科分出而独立成科。我们的研究结果支持Smith et al.（2006）对鳞毛蕨科的重新界定，但被作者暂时置于鳞毛蕨科的三个属：大膜盖蕨属Leucostegia、肿足蕨属Hypodematium和Didymochlaena应该从鳞毛蕨科分立出去;红腺蕨属Diacalpe、毛枝蕨属Leptorumohra和黔蕨属Phanerophlebiopsis应该作为鳞毛蕨科的成员，同时被Smith et al.（2006）保留在叉蕨科的黄腺羽蕨属Pleocnemia也应该作为鳞毛蕨科成员。鳞毛蕨科下分为四个主要的分支：鳞毛蕨支dryopteroids、耳蕨支polystichoids、肋毛蕨支ctenitoids和舌蕨支elaphoglossoids。鳞毛蕨支和耳蕨支互为姐妹群，舌蕨支是其他三个分支的姐妹群。 玉龙蕨属Sorolepidium、柳叶蕨属Cyrtogonellum和鞭叶蕨属Cyrtomidictyum是鳞毛蕨科中的三个东亚特有属，这三个特有属的分类等级和系统位置在不同的分类系统中存在争议。本文对rbcL基因进行分析并结合孢子扫描电镜观察，不支持玉龙蕨属成为一个独立的属，而应该作为耳蕨属的异名。利用rbcL、atpB、trnL-F和rps4-trnS四个DNA片段对柳叶蕨属和鞭叶蕨属进行的系统学分析，支持鞭叶蕨属作为一个独立的属，并且位于整个耳蕨类植物的基部位置。柳叶蕨属同耳蕨属近缘，尤其是同耳蕨属的细裂耳蕨组Sphaenopolystichum、半开羽耳蕨组Haplopolystichum和戟叶耳蕨组Crucifilix关系较近。但是柳叶蕨属的分类等级以及与耳蕨属的属间界限尚需要进一步研究。 2.叉蕨科的分子系统学研究 对rbcL和atpB两个基因片段的单独和联合分析均表明，秦仁昌定义的叉蕨科Tectariaceae不是一个自然的单系类群。在系统发育树上，肋毛蕨属Ctenitis、轴鳞蕨属Dryopsis、节毛蕨属Lastreopsis和黄腺羽蕨属Pleocnemia与鳞毛蕨科聚在一起构成一个强支持的分支。当把上述四个属排除以后，叉蕨属Tectaria、轴鳞蕨属Ctenitopsis、地耳蕨属Quercifilix、牙蕨属Pteridrys和沙皮蕨属Hemigramma形成一个单系类群，并得到很好的支持，该单系类群同条蕨科、骨碎补科和水龙骨科形成姐妹群关系。该单系类群同目前Smith et al.（2006）对叉蕨科的定义一致。在rbcL基因单独分析中，爬树蕨属Arthropteris同叉蕨属－沙皮蕨属聚在一起，但支持率较低。 3.藤蕨科的分子系统学研究及拟贯众属的系统位置 根据对薄囊蕨类114个分类群的rbcL基因和30个代表类群的rbcL、atpB和accD基因的系统发育分析，发现传统的藤蕨科Lomariopsidaceae不是单系类群，除了藤蕨属和Thysanosoria仍然为藤蕨科成员外，藤蕨科的主要成员（实蕨属Bolbitis、网藤蕨属Lomagramma、舌蕨属Elaphoglossum和Teratophyllum）同鳞毛蕨科植物聚在一起，因此应该被归并到鳞毛蕨科。根据Smith et al.（2006）对藤蕨科的最新定义，藤蕨科包括藤蕨属Lomariopsis、肾蕨属Nephrolepis等在内的4个属。但是本文的研究不支持把肾蕨属作为藤蕨科成员，而应该作为一个独立的分类单元，即成立肾蕨科更为合适。根据我们的分析，拟贯众属Cyclopeltis既不是鳞毛蕨科也不是叉蕨科成员，而与藤蕨属Lomariopsis聚成一个强支持的姐妹群。叶片奇数一回羽状、侧生羽片以关节着生于叶轴，叶脉游离等形态特征支持两者的近缘关系。

Limited genetic structure of Gulf Menhaden (Brevoortia patronus), as revealed by microsatellite markers developed for the genus Brevoortia (Clupeidae)

Long-term sustainable management of wild populations should be based on management actions that account for the genetic structure among populations. Knowledge of genetic structure and of the degree of demographic exchange between discreet [sic] populations allows managers to better define management units. However, adequate gene loci for population assessments are not always available. In this study, variable co-dominant DNA loci in the heavily exploited marine genus Brevoortia were developed with a microsatellite-enriched DNA library for the Gulf Menhaden (Brevoortia patronus). Microsatellite marker discovery was followed by genetic characterization of 4 endemic North American Brevoortia species, by using 14 novel loci as well as 5 previously described loci. Power analysis of these loci for use in species identification and genetic stock structure was used to assess their potential to improve the stock definition in the menhaden fishery of the Gulf of Mexico. These loci could be used to reliably identify menhaden species in the Gulf of Mexico with an estimated error rate of α=0.0001. Similarly, a power analysis completed on the basis of observed allele frequencies in Gulf Menhaden indicated that these markers can be used to detect very small levels of genetic divergence (Fst≈0.004) among simulated populations, with sample sizes as small as n=50 individuals. A cursory analysis of genetic structure among Gulf Menhaden sampled throughout the Gulf of Mexico indicated limited genetic structure among sampling locations, although the available sampling did not reach the target number (n=50) necessary to detect minimal values of significant structure.

Predicting the distribution of Vibrio vulnificus in Chesapeake Bay

Vibrio vulnificus is a gram-negative pathogenic bacterium endemic to coastal waters worldwide, and a leading cause of seafood related mortality. Because of human health concerns, understanding the ecology of the species and potentially predicting its distribution is of great importance. We evaluated and applied a previously published qPCR assay to water samples (n = 235) collected from the main-stem of the Chesapeake Bay (2007 – 2008) by Maryland and Virginia State water quality monitoring programs. Results confirmed strong relationships between the likelihood of Vibrio vulnificus presence and both temperature and salinity that were used to develop a logistic regression model. The habitat model demonstrated a high degree of concordance (93%), and robustness as subsequent bootstrapping (n=1000) did not change model output (P > 0.05). We forced this empirical habitat model with temperature and salinity predictions generated by a regional hydrodynamic modeling system to demonstrate its utility in future pathogen forecasting efforts in the Chesapeake Bay.

Larvae of Dactylopsaron dimorphicum (Perciformes: Percophidae) from oceanic islands in the southeast Pacific

Percophids are a family of small marine benthic fishes common over soft bottoms from inshore to the outer slopes in tropical to temperate regions of the Atlantic and in the Indo-West and southeast Pacific (Reader and Neira, 1998; Okiyama, 2000). Five species belonging to four genera have been recorded around the Salas y Gómez Ridge in the southeast Pacific, all of which are endemic to the area except for Chrionema chryseres, a species which also occurs off the Hawaiian Islands and Japan (Parin, 1985, 1990; Parin et al., 1997). Of these five species, larval stages have been described only for Osopsaron karlik and Chrionema pallidum (Belyanina 1989, 1990).

横断山特有植物区系的物种多样性与生物地理学研究

前人对横断山区特有植物的宏生态学研究已有一定的资料积累，对横断山区在全球生物多样性保护中的关键地位和特有现象已有基本共识：（1）横断山区是全球生物多样性关键地区和生物多样性优先重点保护的热点地区之一；（2）横断山区不仅是中国的三大特有植物分布中心之一，而且是新特有中心和物种分化中心，特有现象的生态成因大于历史成因。横断山区种子植物种类丰富，且具有较高的特有现象，其中既有古特有成分也有新特有成分但更多的是新特有成分。但目前，关于横断山区特有现象与生态特征、特有种分布与物种丰富度、特有种丰富度与海拔梯度以及植被的关系等仍缺乏深入的研究。针对上述问题，本论文初步研究了横断山区（1）特有种的生活型、种子散布方式、传粉方式和繁育系统等生态特征，各生态特征之间的相关关系，以及它们的进化地位；（2）特有种的水平分布格局及环境成因；和（3）特有种和生态特征沿海拔梯度的变化规律。主要结论如下： 1．横断山区含特有植物2396个分类群（包括种、亚种和变种），隶属于372属90科。特有类群在372个属中的分布非常不均匀，约一半的特有类群（50.2%）集中分布在较少的属中（占总属数的6%），含特有类群的属大小差异很大，重要的大属是：马先蒿属、乌头属、杜鹃花属、翠雀属、紫堇属、虎耳草属、报春花属、小檗属、黄芪属、凤毛菊属、蚤缀属、橐吾属、龙胆属、景天属、柳属、苔草属、香茶菜属、凤仙花属、紫菀属、鼠尾草属和蝇子草属等，这些属多以横断山为分布和分化中心，为新特有成分。另外，地面芽植物和隐芽植物等占有绝对优势，而矮高位芽植物、高位芽植物、地面芽植物等占有较低的比例，反映了该特有区系的高山、亚高山植物区系特点，但又不乏与热带植物区系的渊源。与世界其它高山地区相比，该特有区系中的地面芽植物、隐芽植物和一年生植物有较高的比例。 2．草本、风力散布种子、昆虫传粉和两性花等为该特有植物区系的优势生态特征。除传粉方式和生活型间无明显的相互作用外，其它生态特征间都存在明显的正相关关系。定量化研究结果表明该特有区系特有种的传粉方式进化可塑性很小，处于保守（或原始）的进化地位。相反，种子散布方式和繁育系统的进化可塑性较大，处于衍生（或进化）的进化地位，是不同种系在历史发育过程中不同阶段的产物。生活型的进化地位在种间有较大的分异，具较小或较大可塑性的各占有一定的比例，因此生活型中既保留处于保守（或原始）的进化地位也有处于衍生（或进化）的进化地位，两种进化状态的生活型在该特有区系中并存，该特征不但是种系在进化早期决定的，同时也是不同种系在进化过程中获得的。 3． 特有种的分布不均匀，较高比例的特有种局限分布在较少的几个地区，北纬28-29°线是特有种丰富度重要的南北分界线。聚类分析结果表明其分布可以划分为3个物种聚集群：（1）北纬28-29°以北、北纬34°以南的藏东川西北物种聚集群；（2）北纬28-29°以南、北纬26°以北的藏东南—滇西北—川西南物种聚集群；和（3）北纬26°以南、北纬25°以北的滇西北物种聚集群。其中，藏东南—滇西北—川西南物种聚集群特有种丰富度最高，是横断山区特有种分布的核心区；滇西北物种聚集群特有种丰富度最低，藏东川西北物种聚集群特有种丰富度居中。藏东南—滇西北—川西南物种聚集群之所以特有种类丰富很有可能与该小区具有高的物种丰富度和落叶阔叶林、常绿/落叶针叶林的分布有密切的联系。在区域尺度上，横断山区总物种的分布中心与特有中心存在分异。尽管两者都处于北纬28-29°以南、北纬26°以北的藏东南—滇西北—川西南小区，但特有中心具有较小的分布范围，而总物种的丰富度中心分布范围较大。因此，我们推测特有分布中心不但与落叶阔叶林和常绿/落叶针叶林有密切的联系，而且也与物种丰富度有关。特有种与植被类型尤其是与常绿阔叶林和落叶阔叶林的关系研究对揭示特有现象的发生、性质和特点有一定的意义，有关研究尚需进一步开展。此外，海拔高差、区系物种丰富度、地理位置和单元面积为特有种丰富度地理分布的总体变异提供了74.7%的解释。 4．研究地区的南缘和东南缘有高比例的木本特有植物，而草本特有植物的比例相对最低，反映出该横断山区南缘的特有植物与热带植物区系的渊源关系，其成分多为一些古老的特有成分；而高比例和较高比例的草本特有植物在研究地区普遍存在，包括研究区域南、北的大部分地区，显然草本特有植物是横断山的优势特有成分。这种以草本特有植物为特征的高山、亚高山特有植物区系有别于我国其它两大特有植物分布中心。该特有中心的成因的最终解释还需要从物种形成（speciation）、灭绝（extinction）和扩散（biogeographic dispersal）——这三个直接影响一个地区物种数量变化的过程来考虑。 5．特有种丰富度沿海拔梯度呈“单峰” 变化曲线，在中海拔段达到峰值。与中国特有种、世界广布种相比，特有种的峰值最高，最适海拔分布范围最窄。特有率随海拔上升而逐渐增高，但在海拔5000-5500 m左右突然上升，在海拔6000 m左右达到最高值。说明（1）物种分布区和丰富度存在着正相关，即特有种具有小的分布区，而广布种的分布区较大；（2）特有率的线性变化规律反映了物种丰富度和特有现象之间的关系，即物种丰富度最大峰值以上海拔段物种数量减少，但特有率增加，说明了该特有区系的高海拔性质。 6．特有种沿海拔梯度表现出大小明显不同的物种聚集群，可能与横断山区的山地垂直气候及相应的植被类型有关。在2600-4600 m海拔段特有种数量最多，这可能与该海拔段分布着落叶阔叶林和常绿/落叶针叶林有密切的联系。特有植物沿海拔梯度可分为四个不同海拔段的物种聚集群：（1）200-1000 m；（2）1000-2600 m； （3）2600-4600 m；和（4）4600-6400 m。在对应海拔段依次分布着干热河谷稀树灌草丛、常绿阔叶林、落叶阔叶林和常绿/落叶针叶林、高山流石滩稀疏植丛。其中，分布在2600-4600 m海拔段的物种丰富度最高，1000-2600 m海拔段的物种丰富度次之，200-1000 m、4600-6400 m两个海拔段物种丰富度较低。 7．特有区系中占优势的生态特征包括草本、风力散布种子、虫媒传粉、两性花和克隆性等对海拔梯度的适应也呈“单峰” 变化曲线，但该主要特征与次要的生态特征（木本、动物散布种子、风媒传粉、单性花和非克隆性）相比，均具有较高的海拔峰值和较窄的最适分布范围，反映了主要生态特征对中、高海拔的适应特点。主要的生态特征与次要的生态特征并存构成了横断山区特有区系的基本生态特征。

濒危植物大花黄牡丹种子休眠与萌发特性研究

本文以中国极危种大花黄牡丹种子为研究材料，对其种子生物学、休眠与萌发特性进行研究，采用变温层积和激素处理解除种子休眠，并通过生物抑制物测试、胚培养、激素含量动态变化的测定，研究种子休眠的原因，从根本上解决了大花黄牡丹迁地保护过程中种子繁殖的技术难题，并初步探讨了大花黄牡丹致濒因素与种子休眠及萌发特性的关系，结果表明： 1.大花黄牡丹种子饱实度高，活力强，种皮较坚硬，但不存在吸水障碍，干燥条件下易因失水而丧失活力。迁地保护冷室中盆播18个月方可出苗，出苗率约4%;变温层积实验表明该种具有典型的下胚轴休眠和上胚轴生长抑制。 2.胚组织培养表明：带胚乳和种皮的胚不能萌发，剥除种皮和胚乳后，胚根可萌发，胚芽不生长，GA3处理上胚轴可促进生长，说明种皮和胚乳是导致大花黄牡丹种子下胚轴休眠的关键因素，而上胚轴生长抑制与胚本身关系更密切。 3.抑制物质提取实验表明：大花黄牡丹种子胚乳、种皮、胚等各部位的浸提液均存在抑制小白菜种子萌发的物质，且抑制作用依次增强，说明胚乳和胚是其下胚轴休眠的主要原因。经过暖层积（15 ℃/90 d）种子（未解除上胚轴生长抑制）的胚根、子叶、胚轴浸提液对小白菜和已解除休眠的大花黄牡丹种子的萌发均有不同程度的抑制作用，并依次减弱，说明种胚本身的抑制物质是导致生理休眠的主要原因。 4.变温层积处理和外源激素实验表明：新鲜种子采收后15 ℃暖层积3个月生根率可达85%，下胚轴休眠解除对温度要求严格，高于或低于15 ℃及变温条件均不利于下胚轴萌发;暖层积90d、根长大于6 cm种子再经过60～80 d/ 5 ℃冷层积，即可有效解除大花黄牡丹种子上胚轴的生长抑制，出芽率达80%，最终出苗率68%。不同浓度GA3及不同处理时间促进上胚轴伸长实验结果显示，GA3　400 mg/L浸种根长大于1.5 cm的种子2 h，出芽率可达100%，可以完全解除上胚轴的生长抑制作用。 5.休眠萌发过程中种子各部位内源激素含量动态变化分析结果说明，初始状态脱落酸含量高是导致大花黄牡丹种子下胚轴休眠、上胚轴生长抑制的主要原因之一，且上胚轴抑制与子叶、下胚轴和根的脱落酸含量密切相关;同时认为种子各部位初始生长素含量水平低是导致其休眠的另一个主要原因;变温层积过程中胚各部位脱落酸含量的急剧下降和生长素的迅速升高是解除休眠的关键;同时发现赤霉素在解除休眠和促进萌发过程中起着重要的促进作用，外源GA3能够有效地打破上胚轴的深度休眠。玉米素核苷在休眠与萌发进程中变化趋势与生长素和赤霉素相似，说明其对种子胚根和上胚轴的萌发和生长具有一定的促进作用。

Some parasitic Copepoda from Sri Lanka (Ceylon) with a synopsis of parasitic Crustacea from Ceylonese freshwater fishes

This report comprises a summary of parasitic copepods from fishes in Ceylon, as isolated from the branchial material of fishes belonging to previous collections. Seven copepod species are described in detail, as well as one species of Branchiura and one species of Isopoda. Caution is advocated to avoid further introductions via parasite-infested fishes, since only four of the above species are endemic.

Results of the Austrian-Ceylonese hydrobiological mission 1970 of the 1st Zoological Institute of the University of Vienna (Austria) and the Department of Zoology of the Vidyalankara University of Ceylon, Kelaniya. Pt. 7. Dryopidae and Elminthidae of Ceylon

This report comprises a summary of parasitic copepods from fishes in Ceylon, as isolated from the branchial material of fishes belonging to previous collections. Seven copepod species are described in detail, as well as one species of Branchiura and one species of Isopoda. Caution is advocated to avoid further introductions via parasite-infested fishes, since only four of the above species are endemic.

An analysis of records of gillnet fishing in Pilkington Bay, Lake Victoria

Among the commercially valuable species occurring in Lake Victoria the two endemic species of Tilapia, T. esculenta (Graham) and T. variabilis (Boulenger) still provide the basis for the fishery in many areas, though in some places the non-cichlid species are more important, for example in the Kagera river area where large numbers of Labeo victorianus and Schilbe mystus migrate seasonally into the river. Although certain species show these habitat preferences. Most of the coastline of Lake Victoria supports a wide variety of species, all of which contribute to the commercial fishery. Over the past ten years the East African Fisheries Research Organisation has fished experimentally a number of stations within a radius of fifty miles from Jinja. These stations have been selected so as to cover a variety of habitats which range from sheltered bays to exposed coastlines. The records discussed in this paper are the result of fishing operations carried out in Pilkington Bay over the period December 1954 to November 1956. This series of fishing operations was carried out in an attempt to assess the composition of the fish populations contained in this area. To this end a variety of nets of differing mesh sizes were used. Pilkington Bay is about ten square miles in extent and is an irregular and deep indentation on the north coast of Buvuma Island. It lies within the sleeping siclmess area where fishing by Africans is prohibited. When selecting an experimental fishing ground, it is important to have some idea of the amount of commercial fishing being carried out in the area. Throughout the period of this experiment a few African fishermen were seen operating illegally in the area, but it is unlikely that their efforts greatly affected the results obtained. Pilkington Bay was fished by E.A.F.R.O. in 1949 and 1950 (Lowe McConnell 1956) but since then till the present experiment, fishing in the area has been infrequent.

The marine linefish resources of Moçambique (status, development and future research)

Marine line-fishing is seen to be of major importance to the social well-being and economy of many coastal people of Mozambique. Aspects of the artisanal and semi-industrial fisheries of Mozambique are described and a recent significant increase in effort is noted. Landings are seen to comprise a high proportion of vulnerable, endemic species, several of which are shared with neighbouring South Africa. Trends in CPUE, sex ratios and yield per recruit suggest that future landings may decline if conservative management is not introduced. Strategies for data collection and biological research on key species are proposed.

Embryonic stages and eye-specific gene expression of the local cyprinoid fish Anabarilius grahami in Fuxian Lake, China

Anabarilius grahami is a cyprinoid fish endemic to Fuxian Lake, Yunnan, China. In this study, a comprehensive staging series of A. grahami was produced. The embryonic development of A. grahami was divided into six main periods: zygote period, cleavage per

Molecular phylogenetic systematics of twelve species of acipenseriformes based on mtDNA ND4L-ND4 gene sequence analysis

Acipenseriformes is an endangered primitive fish group, which occupies a special place in the history of ideas concerning fish evolution, even in vertebrate evolution. However, the classification and evolution of the fishes have been debated. The mitochondrial DNA (mtDNA) ND4L and partial ND4 genes were first sequenced in twelve species of the order Acipenseriformes, including endemic Chinese species. The following points were drawn from DNA sequences analysis: (i) the two species of Huso can be ascribed to Acipenser; (ii) A. dabryanus is the mostly closely related to A. sinensis, and most likely the landlocked form of A. sinensis; (iii) genus Acipenser in trans-Pacific region might have a common origin; (iv) mtDNA ND4L and ND4 genes are the ideal genetic markers for phylogenetic analysis of the order Acipenseriformes.