Lothar Krienitz

Contribution of hot spring cyanobacteria to the mysterious deaths of Lesser Flamingos at Lake Bogoria, Kenya

Cyanobacterial mats at hot springs on the shore of the alkaline Lake Bogoria, Kenya, were investigated regarding species community and cyanobacterial toxin content. The hepatotoxins microcystin-LR, -RR, -LF and -YR, and the neurotoxin anatoxin-a were present. The mats were dominated by Phormidium terebriformis, Oscillatoria willei, Spirulina subsalsa and Synechococcus bigranulatus. The concentration of microcystins in mat samples, ranged from 221 to 845 microg microcystin-LR equivalents g(-1) DW of mat. Anatoxin-a concentrations ranged from 10 to 18 microg g(-1) DW of mat. A contribution of the cyanobacterial toxins from the hot spring mats to the mass mortalities of Lesser Flamingos is suggested by: (a), the presence of hot spring cyanobacterial cells and cell fragments, and high concentrations of the cyanobacterial hepato- and neurotoxins in flamingo stomach contents and faecal pellets; (b), observations of neurological signs of bird poisoning at the lake. Cyanobacterial toxins in stomach contents, intestine and fecal pellets were 0.196 microg g(-1) fresh weight (FW) for the microcystins and 4.34 microg g(-1) FW for anatoxin-a. Intoxication with cyanobacterial toxins could occur by uptake of detached cyanobacterial cells from the mats, as the flamingos need to drink fresh or brackish water, and to wash their feathers daily, which they do in the vicinity of the hot springs, where salinity is lower than in the main body of water of the lake.

Phylogenetic relationship of Chlorella and Parachlorella nov. gen. (Chlorophyta, Trebouxiophyceae)

Abstract Chlorella is one of the archetypical forms of coccoid green algae and one of the best-studied phototrophic eukaryotes. However, its systematics remains enigmatic due to conflicts between morphological and molecular phylogenetic approaches. The sequences of the 18S ribosomal RNA gene of nine strains of Chlorella and related taxa, and the ITS2 region of 17 strains of Chlorellaceae were determined and included in phylogenetic analyses. The secondary structure of the ITS2 region of C. vulgaris was compared to that of Parachlorella beijerinckii. All phylogenetic analyses showed that the Chlorellaceae form a clade within the Trebouxiophyceae. The Chlorellaceae studied here are divided into two sister groups: (1) the Parachlorella-clade with Parachlorella beijerinckii gen. et sp. nov. and P. kessleri comb. nov. as well as Diclosler acuatus and Closteriopsis acicularis; and (2) the Chlorella-clade including the ‘true’ spherical Chlorella species C. vulgaris, C. lobophora and C.sorokiniana. The latter are intermixed with taxa that differ in morphology and were formerly classified in other families of coccoid green algae: Actinastrum hantzschii (formerly Coelastraceae), Diacanthos belenophorus (formerly Oocystaceae), Dictyosphaerium pulchellum (formerly Botryococcaceae), Didymogenes anomala and D. palatina (formerly Scenedesmaceae) and Micractinium pusilium (formerly Micractiniaceae). It was shown that morphological criteria traditionally used for classification (e.g. spines, mucilaginous envelopes or the formation of colonies or coenobia) are burdened with a high degree of phenotypic plasticity. These plastic traits represent adaptations to environmental factors in order to optimize floating in the water column and to resist grazing pressure. A genus and species concept in Chlorellaceae remains provisional, because, in most cases, there is still a lack of classical and molecular approaches to identify clear phylogenetic lineages.

Present state of the systematics of planktonic coccoid green algae of inland waters

This review discusses the main developments in the systematics of coccoid green algae over the last three decades. The relationships of key groups of planktonic coccoid green algae are shown in the phylogenetic trees of Chlorophyceae and Trebouxiophyceae. The trees clearly show that the morphology of these algae do not adequately reflect their phylogenetic position. Different phylogenetic species can be hidden under one and the same morphotype. As most of the genera have a polyphyletic origin, they are in need of a systematic re-evaluation. Species classification using the phylogenetic species concept resulted in the establishment of new genera with smaller numbers of species and the description of new species that are not distinguishable by light microscopy. An overview of genera is given in tables and the revised designations of species as contained in the harmonized taxon list of the European Water Framework Directive lists is provided. In this transitional phase from an artificial to a more natural systematics of algae, field biologists and ecologists as well as molecular biologists must strengthen their interdisciplinary cooperation. The alignment of eco-functional groups of algae with true species identities using the barcoding conception will provide a better understanding of the interaction between organisms and their environment.

Deep-layer autotrophic picoplankton maximum in the oligotrophic Lake Stechlin, Germany: origin, activity, development and erosion

Autotrophic picoplankton (APP) abundance, primary production and vertical distribution were studied in the oligotrophic Lake Stechlin (northeastern Germany) in 1994 and 1995. Within the euphotic zone APP contributed 34% of phytoplankton biomass and 35% of primary production. Annual average APP cell number was 209 × 103 cells ml−1, of which 95% were unicellular cyanobacteria, 2% were colonial cyanobacteria and 3% were eukaryotes. Three ecologically and/or morphologically different groups of APP were recognized: (i) unicellular cyanobacteria belonging to the genus Cyanobium, (ii) eukaryotic species growing in early spring under isothermal conditions and (iii) cyanobacteria, partly colonial species, growing in the stratified period in the euphotic zone. Three species of eukaryotic green algae were identified: Choricystis minor, Neocystis diplococca and Pseudodictyosphaerium jurisii, the latter two being colonial. This is the first record of the occurrence of colonial eukaryotes potentially of APP size in fre...

PHYLOGENY OF THE HYDRODICTYACEAE (CHLOROPHYCEAE): INFERENCES FROM rDNA DATA1

The hydrodictyacean green algal lineage has been the focus of much research due to the fossil record of at least some members, their ornamented cell walls, and their distinctive reproductive strategies. The phylogeny of the family was, until recently, exclusively morphology based. This investigation examines hydrodictyacean isolates from several culture collections, focusing on sequences from ribosomal data: 18S rDNA, 26S rDNA (partial), and internal transcribed spacer (ITS)‐2 data. Results from phylogenetic analyses of independent and combined data matrices support the Hydrodictyaceae as a monophyletic lineage that includes isolates of Chlorotetraedron, Hydrodictyon, Pediastrum, Sorastrum, and Tetraedron. Phylogenetic analyses of rDNA data indicate that the three‐dimensional coenobium of Hydrodictyon is evolutionarily distinct from the three‐dimensional coenobium of Sorastrum. The more robust aspects of the ITS‐2 data corroborate the 18S+26S rDNA topology and provide a structural autapomorphy for the Hydrodictyaceae and Neochloridaceae, that is, an abridgment of helix IV in the secondary structure. The rDNA data do not support monophyly of Pediastrum but rather suggest the existence of four additional hydrodictyacean genera: Monactinus, Parapediastrum, Pseudopediastrum, and Stauridium.

Hyaloraphidium curvatum is not a green alga, but a lower fungus; Amoebidium parasiticum is not a fungus, but a member of the DRIPs.

The unicellular heterotrophic protist Hyaloraphidium is classified with a family of green algae, the Ankistrodesmaceae. The only species that exists in pure culture and that is available for taxonomic studies is H. curvatum. Comparative 18S ribosomal RNA sequence analyses showed that H. curvatum belongs to the fungi rather than to the algae. Within the fungi, H. curvatum preferentially clustered with Chytridiomycetes. Unlike Chytridiomycetes, H. curvatum propagates by autosporulation, and the presence of flagella has never been reported. Transmission electron microscopy indicated that H. curvatum in some respects resembles Chytridiomycetes, but no elements of a flagellar apparatus were detected. The habitus of H. curvatum is unlike that of other fungi except the trichomycete Amoebidium parasiticum. The cell wall sugar composition of H. curvatum was unique, but to some extent resembled that of A. parasiticum. However, H. curvatum and A. parasiticum are not closely related to each other according to 18S rRNA sequence data. Moreover, A. parasiticum clustered with protistan animals, the Mesomycetozoa (DRIPs). Combined molecular, ultrastructural and chemical data do not allow assignment of H. curvatum to any recognized clade of fungi. This suggests that H. curvatum may represent an independent evolutionary lineage within the fungi.

TRADITIONAL GENERIC CONCEPTS VERSUS 18S rRNA GENE PHYLOGENY IN THE GREEN ALGAL FAMILY SELENASTRACEAE (CHLOROPHYCEAE, CHLOROPHYTA)

Coccoid green algae of the Selenastraceae were investigated by means of light microscopy, TEM, and 18S rRNA analyses to evaluate the generic concept in this family. Phylogenetic trees inferred from the 18S rRNA gene sequences showed that the studied species of autosporic Selenastraceae formed a well‐resolved monophyletic clade within the DO group of Chlorophyceae. Several morphological characteristics that are traditionally used as generic features were investigated, especially the arrangement of autospores in the mother cells, colony formation, and pyrenoid structure. The parallel arrangement of autospores was confirmed for the genera Ankistrodesmus, Podohedriella, and Quadrigula. In mother cells of Monoraphidium and Kirchneriella, the autospores were arranged serially. Colony formation was either stable (Quadrigula) or variable (Ankistrodesmus, Podohedriella) within genera. All strains studied possessed naked or starch‐covered pyrenoids within the chloroplast. The pyrenoid matrix was homogenous or penetrated by thylakoids. In contrast to considerations of traditional systematics, the present study showed that the presence and structure of pyrenoids are unsuitable for differentiation of genera in Selenastraceae. Furthermore, the molecular analyses showed that any morphological criterion considered so far is not significant for the systematics of the Selenastraceae on the generic level. Species assigned to different genera such as Ankistrodesmus and Monoraphidium were not monophyletic and therefore not distinguishable as separate genera. Species of Monoraphidium appeared in four different lineages of the Selenastraceae. Our phylogenetic analyses support earlier discussions to abandon the common practice of conceiving “small” genera (i.e. genera that are differentiated from other genera by only a few diacritic characteristics and that contain only a small number of species) and to reestablish “large” genera of Selenastraceae such as Ankistrodesmus.

Fluctuating algal food populations and the occurrence of lesser flamingos (Phoeniconaias minor) in three Kenyan rift valley lakes.

The last two decades have witnessed increasing episodes of lesser flamingo die‐offs in East Africa. Based on data on phytoplankton composition, biomass, and flamingo population density in three alkaline‐saline lakes of Kenya (Bogoria, Nakuru, and Oloidien) in 2001–2010, this study explored the link between sudden flamingo deaths and fluctuations in algal food quantity and quality. The phytoplankton biomass ranged from 13 to 768 mg · L−1. Similarly, flamingo numbers varied widely from <1,000 to >500,000 individuals in the study lakes. The dominance of the cyanobacterium Arthrospira fusiformis (Woron.) Komárek et J. W. G. Lund was interrupted at irregular intervals in each lake and replaced partly by populations of different species of the nostocalean Anabaenopsis or by the picoplanktonic chlorophyte Picocystis salinarum Lewin. The populations of Anabaenopsis have the potential of blocking the flamingo food filtration system with their large and slimy colonies; moreover, they are able to produce cyanotoxins. Estimates of flamingo populations suggest that low flamingo numbers coincided with periods of low algal food quantity and/or poor quality. A food deficit can be theorized to have two effects on the flamingos: (i) it weakens them to the point of being susceptible to attacks of infective diseases, such as the ones caused by Mycobacterium avium and Pseudomonas aeruginosa, and (ii) it predisposes them to poisoning by cyanotoxins and pollutants, by reducing their capacity to handle toxic substances. This study therefore concludes that the challenges facing the flamingos are associated with changes in their environment, which affect food and water supply.

Cyanobacterial toxins in Lake Baringo, Kenya

Abstract This paper presents data on the first identification, characterization and quantification of hepatotoxic microcystins and neurotoxic anatoxin-a in water samples of Lake Baringo, Kenya. The shallow turbid Lake Baringo was investigated five times between June 2001 and May 2002. The phytoplankton community was mainly dominated by the cyanobacterium Microcystis aeruginosa . Due to the high turbidity the phytoplankton biomass was low, ranging between 1.5 and 8.2 mg L −1 . High mean total phosphorus concentration (1.0 mg L −1 ) and mean total nitrogen concentration (2.8 mg L −1 ) typical for hypertrophic lakes were found. Using HPLC technique the hepatotoxins microcystin-LR, -RR and -YR and the neurotoxin anatoxin-a were detected in the water samples. The microcystin concentrations varied from 310 to 19800 μg microcystin-LR equivalents g −1 DW and the anatoxin-a concentration ranged from 270 to 1260 μg g −1 DW. To our knowledge this is the first evidence of cyanobacterial toxins in Lake Baringo.

ITS2 sequence-structure phylogeny in the Scenedesmaceae with special reference to Coelastrum (Chlorophyta, Chlorophyceae), including the new genera Comasiella and Pectinodesmus

Hegewald E., Wolf M., Keller A., Friedl T. and Krienitz L. 2010. ITS2 sequence-structure phylogeny in the Scenedesmaceae with special reference on Coelastrum (Chlorophyta, Chlorophyceae). Phycologia 49: 325–335. DOI: 10.2216/09-61.1 Sequences and secondary structures of the nuclear-encoded internal transcribed spacer 2 (ITS2) ribosomal RNA of nine Coelastrum taxa, Asterarcys quadricellulare (Coelastraceae), Westella botryoides (hitherto Oocystaceae) and Dimorphococcus lunatus (Scenedesmaceae) were determined and compared with existing GenBank entries of scenedesmacean taxa (Desmodesmus, Enallax, Neodesmus, Scenedesmus). Phylogenetic analyses showed that the studied Coelastrum taxa belong to several different lineages within the Scenedesmaceae: five Coelastrum taxa (Coelastrum microporum, Coelastrum astroideum, C. astroideum var. rugosum  =  Coelastrum rugosum, Coelastrum pseudomicroporum and Coelastrum sphaericum incl. Coelastrum proboscideum) form monophyletic clades, whereas two strains labeled Coelastrum morum belong to different genera. The African strain of C. morum clusters with Coelastrum cambricum. The Finnish strain labeled C. morum clusters with Asterarcys, Dimorphococcus and Hariotina. According to its morphology this strain belongs to Coelastrella, related to Coelastrella saiponensis. Westella botryoides belongs to a separate clade within the Scenedesmaceae. Coelastrum reticulatum is positioned in the clade with Asterarcys, Dimorphococcus and Coelastrella; hence its separation in a separate genus, as originally described (Hariotina), is justified. In general, the phylogenetic analysis of ITS2 data shows that the Coelastraceae are included in the monophyletic Scenedesmaceae, and thus the splitting into two families is not justified, but they belong to the monophyletic subfamily Coelastroidea. The genera Comasiella and Pectinodesmus are newly erected, and several new combinations are proposed.