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Improved Methods for Bulk Cultivation and Fixation of Loxodes Ciliates for Fluorescence Microscopy 荧光显微镜下纤毛虫批量培养和固定的改进方法
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.1016/j.protis.2022.125905
Brandon Kwee Boon Seah, Christiane Emmerich, Aditi Singh, Estienne Carl Swart

Loxodes is one of the best ecologically characterized ciliate genera with numerous intriguing physiological abilities, including gravity-sensing organelles and nitrate respiration. However, these cells have been considered challenging to cultivate in bulk, and are poorly preserved by conventional fixatives used for fluorescence microscopy. Here we describe methods to grow and harvest Loxodes cells in bulk with liquid soil extract medium, as well as a new fixative called ZFAE (zinc sulfate, formaldehyde, acetic acid, ethanol) that can fix Loxodes cells more effectively than buffered formaldehyde or methanol. We show that ZFAE is compatible with immunofluorescence and the nuclear stain DAPI. Loxodes is thus now amenable to long-term maintenance, large-scale growth, and modern cell biology investigations of monoclonal strains in laboratory conditions.

Loxodes是最具生态学特征的纤毛虫属之一,具有许多有趣的生理能力,包括重力感应细胞器和硝酸盐呼吸。然而,这些细胞被认为难以批量培养,并且用荧光显微镜使用的常规固定液保存得很差。在这里,我们描述了用液体土壤提取物培养基大量生长和收获Loxodes细胞的方法,以及一种名为ZFAE(硫酸锌、甲醛、乙酸、乙醇)的新型固定剂,它比缓冲的甲醛或甲醇更有效地固定Loxodes细胞。我们发现ZFAE与免疫荧光和核染色DAPI兼容。因此,Loxodes现在可以在实验室条件下对单克隆菌株进行长期维护,大规模生长和现代细胞生物学研究。
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引用次数: 0
High Environmentally Induced Plasticity in Spore Size and Numbers of Nuclei per Spore in Physarum albescens (Myxomycetes) 白色绒泡菌孢子大小和每孢子核数的高环境诱导可塑性
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.1016/j.protis.2022.125904
Jan Woyzichovski , Oleg N. Shchepin , Martin Schnittler

Spore size enables dispersal in plasmodial slime molds (Myxomycetes) and is an important taxonomic character. We recorded size and the number of nuclei per spore for 39 specimens (colonies of 50–1000 sporocarps) of the nivicolous myxomycete Physarum albescens, a morphologically defined taxon with several biological species. For each colony, three sporocarps were analyzed from the same spore mount under brightfield and DAPI-fluorescence, recording ca. 14,000 spores per item. Diagrams for spore size distribution showed narrow peaks of mostly uninucleate spores. Size was highly variable within morphospecies (10.6–13.5 µm, 11–13%), biospecies (3–13%), even within spatially separated colonies of one clone (ca. 8%); but fairly constant for a colony (mean variation 0.4 µm, ca. 1.5%). ANOVA explains most of this variation by the factor locality (within all colonies: 32.7%; within a region: 21.4%), less by biospecies (13.5%), whereas the contribution of intra-colony variation was negligible (<0.1%). Two rare aberrations occur: 1) multinucleate spores and 2) oversized spores with a double or triple volume of normal spores. Both are not related to each other or limited to certain biospecies. Spore size shows high phenotypic plasticity, but the low variation within a colony points to a strong genetic background.

孢子大小决定了黏菌在黏菌中的扩散,是一个重要的分类学特征。本文记录了39个(50 ~ 1000个孢子果皮的菌落)嗜烟黏菌白绒泡菌(Physarum albescens)的大小和每孢子核数。对每个菌落,在明场和dapi荧光下分析同一孢子座上的3个孢子实,每项记录约14000个孢子。孢子大小分布图显示,单核孢子多呈窄峰分布。形态种(10.6-13.5µm, 11-13%)和生物种(3-13%)之间的大小变化很大,即使在一个无性系的空间分离菌落内(约8%);但对于一个菌落来说是相当恒定的(平均变化0.4µm,约1.5%)。方差分析(ANOVA)解释了这种变化的大部分因素地方性(在所有菌落中:32.7%;在同一区域内:21.4%),不同生物物种的差异较小(13.5%),而群落内变异的贡献可以忽略不计(0.1%)。出现两种罕见的异常现象:1)多核孢子和2)超大孢子,体积是正常孢子的两倍或三倍。两者彼此没有关系,也不局限于某些生物物种。孢子大小表现出高度的表型可塑性,但在一个菌落内的低变异表明有很强的遗传背景。
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引用次数: 0
Bland James Finlay (March 16, 1952–December 24, 2021) 布兰德·詹姆斯·芬利(1952年3月16日- 2021年12月24日)
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.1016/j.protis.2022.125906
Tom Fenchel
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引用次数: 0
Significance of the Nudiform and Tectiform Modes of Silica Deposition, Lorica Assembly and Cell Division in Choanoflagellates as Exemplified by Stephanoeca diplocostata and Enibas spp. 以Stephanoeca diplocostata和enspp为例的毛鞭毛虫裸形和隐形二氧化硅沉积、Lorica组装和细胞分裂模式的意义
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.2139/ssrn.4152857
B. Leadbeater, M. Carr
The deposition of silicified costal strips and lorica assembly in choanoflagellates is precisely linked to the cell cycle. A minority of species undergo nudiform division whereby a loricate cell divides to produce a naked daughter cell that deposits a set of costal strips and then assembles a lorica. Most species undergo tectiform division whereby a parent loricate cell produces a set of costal strips, divides and passes on the stored strips to a daughter cell that immediately assembles a lorica. Many phylogenetic analyses recover nudiform and tectiform species as sister-clades giving the impression that they are distinct evolutionary lineages. However, the tectiform species Stephanoeca diplocostata is capable of undergoing nudiform division and depositing costal strips and assembling a lorica with certain modifications in a nudiform manner. The recent discovery of a new genus, Enibas, comprising species with Stephanoeca-like loricae that undergo nudiform cell division and on phylogenetic analysis occur as a sister group to other nudiform species has drawn attention to whether there are also unique features in lorica construction. A range of Enibas loricae is illustrated and it appears that there are unique features which might be interpreted as being derived from a Stephanoeca-like ancestor.
甲藻中硅化肋带的沉积和懒虫的组装与细胞周期密切相关。少数物种经历裸体分裂,懒猴细胞分裂产生一个裸体的子细胞,该子细胞沉积一组肋带,然后组装懒猴。大多数物种都会经历保护态分裂,通过这种分裂,亲本懒猴细胞会产生一组肋带,分裂并将储存的肋带传递给子细胞,子细胞会立即组装懒猴。许多系统发育分析将裸形和保护形物种恢复为姐妹分支,给人的印象是它们是不同的进化谱系。然而,保护型物种Stephanoeca diplocosta能够进行裸体分裂,沉积肋带,并以裸体方式组装经过某些修饰的懒猴。最近发现了一个新属Enibas,该属由具有类似Stephanoeca的loricae的物种组成,这些物种经历了裸体细胞分裂,并在系统发育分析中作为其他裸体物种的姐妹群出现,这引起了人们对lorica构建是否也有独特特征的关注。图中展示了一系列的埃尼巴斯loricae,似乎有一些独特的特征可以被解释为源自类似Stephanoeca的祖先。
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引用次数: 2
Unravelling the Phylogeny of a Common Intestinal Protist: Intrageneric Diversity of Endolimax 揭示一种常见肠道原生生物的系统发育:Endolimax的属内多样性
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.1016/j.protis.2022.125908
Emma Filtenborg Hocke , Mahwash Jamy , Fabien Burki , C. Graham Clark , Christen Rune Stensvold

Endolimax nana is a common endobiont of the human intestine, but members of the genus have also been reported in non-human hosts and in non-intestinal organs. Limited information is available regarding the genetic diversity of Endolimax, which is necessary to delineate species, host specificity and potential differences in clinical impact on the host. Here, we used cloning of PCR products followed by Sanger sequencing and next-generation PacBio Sequencing to obtain Endolimax-related nuclear ribosomal gene sequences and undertook a phylogenetic analysis to gain additional insight into the taxonomy of Endolimax and related organisms. The new sequences confirmed that E. nana forms a discrete clade within the Archamoebae and is related to Endolimax piscium and Iodamoeba. However, we identified substantial sequence divergence within E. nana and evidence for two distinct clades, which we propose to name E. nana ribosomal lineage 1 and E. nana ribosomal lineage 2. Both of the sequencing approaches applied in the study helped us to improve our understanding of genetic diversity across Endolimax, and it is likely that wider application of next-generation sequencing technologies will facilitate the generation of Endolimax-related DNA sequence data and help complete our understanding of its phylogenetic position and intrageneric diversity.

内毒素是一种常见的人类肠道内生物,但该属的成员也在非人类宿主和非肠道器官中被报道。关于Endolimax基因多样性的信息有限,这是描述物种、宿主特异性和对宿主临床影响的潜在差异所必需的。本研究利用PCR产物克隆、Sanger测序和下一代PacBio测序获得了Endolimax相关核糖体基因序列,并进行了系统发育分析,以进一步了解Endolimax及其相关生物的分类。新的序列证实了E. nana在古阿米巴中形成了一个独立的分支,与Endolimax piscium和Iodamoeba有亲缘关系。然而,我们在e.o nana中发现了大量的序列差异和两个不同分支的证据,我们建议将其命名为e.o nana核糖体谱系1和e.o nana核糖体谱系2。研究中采用的两种测序方法都有助于我们提高对Endolimax遗传多样性的理解,下一代测序技术的广泛应用可能会促进Endolimax相关DNA序列数据的生成,并有助于我们完成对其系统发育位置和基因内多样性的理解。
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引用次数: 1
Integrative Studies on Three Epibiotic Epistylis Species (Protozoa, Ciliophora, Peritrichia) in Lake Weishan Wetland, Northern China, Including the Establishment of a New Species 微山湖湿地3种附着生物Epistylis (Protozoa, Ciliophora, Peritrichia)的综合研究及一个新种的建立
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.1016/j.protis.2022.125909
Tong Wu , Yujie Liu , Yan Liu , Borong Lu , Xiao Cao , Shuang Hao , Saleh A. AL-Farraj , Alan Warren , Tengteng Zhang , Zhe Wang

Three epibiotic Epistylis species, i.e., Epistylis weishanensis sp. nov., Epistylis daphniae Fauré-Fremiet, 1905, and Epistylis pygmaeum (Ehrenberg, 1838) Foissner et al., 1999, were investigated based on their living morphology, infraciliature, and small subunit (SSU) rDNA sequence data. Epistylis weishanensis sp. nov. is characterized by its double-layered peristomial lip, contractile vacuole located on the dorsal wall of the infundibulum, infundibular polykinety 3 (P3) composed of three equal-length rows that terminate above infundibular polykinety 1 (P1), 50–62 silverlines between the peristome and the trochal band, and about 30 silverlines between the trochal band and the scopula. Based on previous and newly obtained data for E. daphniae and E. pygmaeum, improved diagnoses and redescriptions are provided including, for the first time, data on their infraciliature. Phylogenetic analyses reveal that all three species do not group within the major clade of Epistylis, supporting the assertion that the genus Epistylis should be an assemblage of morphospecies and therefore needs to be revised.

对3种表生Epistylis weishanensis sp. nov.、Epistylis daphniae faur - fremiet, 1905和Epistylis pygmaum (Ehrenberg, 1838) Foissner et al, 1999的生活形态、基础设施和小亚单位(SSU) rDNA序列进行了研究。微山Epistylis weishanensis sp. 11 .的特征是:双层的壁周唇,位于底窝背壁的可收缩液泡,底窝多动带3 (P3)由三等长的行组成,终止于底窝多动带1 (P1)之上,壁壁与转子带之间有50-62条银线,转子带与scopula之间约有30条银线。根据水蚤E. daphniae和pygmaum的既往和新获得的资料,提供了改进的诊断和重新描述,包括首次提供了它们的基础设施数据。系统发育分析表明,这三个物种都不属于Epistylis的主要分支,这支持了Epistylis属应该是形态种的集合的主张,因此需要修改。
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引用次数: 5
Illustrating the Invisible: The Role of Drawing in the Career of Bland J. Finlay 阐释无形:绘画在布兰德·j·芬利职业生涯中的作用
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-10-01 DOI: 10.1016/j.protis.2022.125907
Carlos F. Finlay

This is a dedicatory article to the role drawing played in the career of Bland J. Finlay FRS, written by his son. It explores some of the many diagrams Bland produced as part of his research, while reflecting on the broader influence of protozoloogical illustrations in art and architectural history.

这是一篇献给绘画在布兰德·j·芬利FRS职业生涯中所扮演角色的文章,由他的儿子撰写。它探讨了布兰德作为其研究的一部分而制作的许多图表中的一些,同时反映了原生动物学插图在艺术和建筑史上的广泛影响。
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引用次数: 0
Some Personal Memories of Bland J. Finlay and Work on Protists in "Priest Pot". 布兰德·J·芬莱的一些个人回忆和《牧师壶》中的主人公作品。
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-09-01 DOI: 10.2139/ssrn.4208046
U. Berninger
With his scientific curiosity and deep understanding of unicellular organisms, Bland J. Finlay could demonstrate that water bodies as small and innocuous as "Priest Pot" in the English Lake District represent a very broad range of protists and their interactions. From this, not only the field of protistan ecology benefitted greatly, but also my personal career. Enclosed are some scientific and personal memories to say "thank you" to Bland.
凭借对科学的好奇心和对单细胞生物的深入了解,布兰德·J·芬莱可以证明,像英格兰湖区的“牧师壶”这样小而无害的水体代表了非常广泛的原生生物及其相互作用。从中受益匪浅的不仅是原生生态领域,还有我个人的事业。随信附上一些科学和个人的回忆,对布兰德说“谢谢”。
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引用次数: 0
The Extraordinarily Rare Ciliate Legendrea loyezae Fauré-Fremiet, 1908 (Haptoria, Ciliophora). 特别罕见的Ciliate Legendarea loyezae Fauré-Fremiet,1908年(Haptoria,Ciliophora)。
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-09-01 DOI: 10.2139/ssrn.4163417
James Weiss, D. Andreou, G. Esteban
Diverse and dynamic communities of ciliates and other microbes thrive in the natural environment, driving the functioning of aquatic ecosystems. Many microbes are present in very low numbers or are dormant in the 'seedbank', escaping detection in environmental surveys and, consequently, remaining underexplored. Here, we report an extraordinarily rare ciliate that was discovered after persistent exploration of freshwater anoxic sediments - Legendrea loyezae Fauré-Fremiet, 1908, a member of the Family Spathidiidae, Order Haptorida. In this study, we present the sixth account of the ciliate since 1908 and reveal its phylogenetic position with the first 18S rRNA data for the genus. We explain the key morphological features of the species, describing a remarkable behaviour in which the ciliate "shapeshifts'' due to its ability of controlled full extension and retraction of its tube-like tentacles. Our results shed light on the similarity of L. loyezae to another ciliate that was first described as Legendrea bellerophon, later moved under a new genus and named Thysanomorpha bellerophon. We question the validity of this taxonomic decision and, based on morphological characters and tentacle movement, we propose moving T. bellerophon back under Legendrea. This study demonstrates how continued and persistent exploration of natural habitats lead to the discovery of microbial communities and species.
多样而动态的纤毛虫和其他微生物群落在自然环境中茁壮成长,推动着水生生态系统的功能。许多微生物的数量非常少,或者在“种子库”中处于休眠状态,逃避了环境调查的检测,因此仍然没有得到充分的探索。在这里,我们报告了一种非常罕见的纤毛虫,它是在对淡水缺氧沉积物进行持续探索后发现的——Legendrea loyezae faur - fremiet, 1908年,是蛛形纲蛛形科的一员。在这项研究中,我们提出了自1908年以来对纤毛虫的第六次描述,并利用该属的第一个18S rRNA数据揭示了其系统发育位置。我们解释了该物种的关键形态特征,描述了一种显着的行为,其中纤毛虫“变形”是由于其控制其管状触须的完全伸展和收缩的能力。我们的研究结果揭示了L. loyezae与另一种纤毛虫的相似性,该纤毛虫最初被描述为Legendrea bellerophon,后来被归入一个新属并命名为Thysanomorpha bellerophon。我们对这一分类决定的有效性提出质疑,并基于形态特征和触手运动,我们提出将白须鲸移回到legendia下。这项研究证明了对自然栖息地的持续探索如何导致微生物群落和物种的发现。
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引用次数: 3
The Morphology, Ultrastructure and Molecular Phylogeny of a New Soil-Dwelling Kinetoplastid Avlakibodo gracilis gen. et sp. nov. (Neobodonida; Kinetoplastea) 一种新的土栖动质体Avlakibodo gracilis gen. et sp. nov. (Neobodonida)的形态、超微结构和分子系统发育Kinetoplastea)
IF 2.5 3区 生物学 Q4 MICROBIOLOGY Pub Date : 2022-08-01 DOI: 10.1016/j.protis.2022.125885
Artem O. Belyaev , Dmitriy G. Zagumyonnyi , Alexander P. Mylnikov , Denis V. Tikhonenkov

Kinetoplastids represent a stockpile of undiscovered protist diversity. Free-living members of this group have been studied less intensively compared to their important parasitic relatives. We have isolated a new soil-dwelling bacteriotrophic kinetoplastid, which is described here as a new genus and new species, Avlakibodo gracilis gen. et sp. nov. Phylogenetic analysis of 18S rRNA genes showed highly supported sister relationship of this protist with the clade uniting Neobodo borokensis, Neobodo curvifilus, Neobodo saliens, Actuariola framvarensis, some Neobodo designis isolates and several environmental sequences, with high statistical support. We have reconstructed the organization of the microtubular cytoskeleton of A. gracilis and determined the origins of the main bands of microtubules. Characteristic ultrastructural features include cytostome associated microtubules (FAS), cytopharynx associated additional microtubules (CMT), microtubular prism (nemadesm) and three microtubular roots (R1, R2 and R3).

着丝质体代表了未被发现的原生生物多样性的储备。与它们重要的寄生亲戚相比,对这一群体的自由生活成员的研究不那么深入。我们分离到了一个新的土壤细菌营养动胞体(Avlakibodo gracilis gen. et sp. 11),本文将其描述为一个新属新种。系统发育分析表明,该原生生物与Neobodo borokensis、Neobodo curvifilus、Neobodo saliens、Actuariola framvarensis、Neobodo designis的一些分离株和一些环境序列具有高度的统计学支持。我们重建了薄叶草微管细胞骨架的组织结构,并确定了微管主要带的来源。典型的超微结构特征包括细胞壁相关微管(FAS)、细胞咽部相关附加微管(CMT)、微管棱镜(nemadesm)和三个微管根(R1、R2和R3)。
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引用次数: 2
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Protist
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