Phylogeny and morphological evolution of the so‐called bougainvilliids (Hydrozoa,Hydroidolina)

We present phylogenetic analyses (parsimony, maximum likelihood and Bayesian inference) for 69 lineages of anthoathecate hydroids based on 18 morphological characters (12 proposed for the first time) plus mitochondrial (16S and COI) and nuclear (18S and 28S) molecular markers. This study aims to test the monophyly of the present concept of the family Bougainvilliidae, assessing its phylogenetic position within Hydroidolina. Our working hypothesis is used as a context for inferring the evolution of certain morphological characters, focusing on the exoskeleton. Our results shed light on some phylogenetic uncertainties within Hydroidolina, delimiting eight well‐supported linages, viz. Hydroidolina, Siphonophorae, Leptothecata, Aplanulata, Filifera II, Filifera III, Capitata and Pseudothecata taxon novum, the latter supported by four morphological synapomorphies. The monophyly of several families was not supported, viz. Bougainvilliidae, Cordylophoridae, Oceaniidae, Rathkeidae and Pandeidae. Some of the genera typically considered in Bougainvilliidae, including Bougainvillia, fell into the clade Pseudothecata, which is consistently reconstructed as the sister group of Leptothecata. We formally suggest that Dicoryne be removed from Bougainvilliidae and placed in the resurrected family Dicorynidae. The exoskeleton was a key feature in the diversification of Hydroidolina, especially with the transition from the bare hydranth to one completely enveloped within the exoskeleton. In this context, bougainvilliids exhibit several intermediate states in the development of the exosarc. Although the concatenated analysis unravels some interesting hypotheses, taxon sampling is still deficient and therefore more data are necessary for achieving a more complete understanding of the evolution and ecology of bougainvilliids and their allies.     

Preliminary Observations on the Nature,Taxonomic Composition,Distribution, Behaviour and Nutrient Requirements of the “Unusual” Algal “Jellies” of Oguta Lake in Southeastern Nigeria

Preliminary observations were made between October 1982 and May 1983 on the nature, taxonomic composition, spatial distribution, buoyancy behaviour and nutrient requirements of some unusual planktonic algal “jellies” in Oguta Lake in Southeastern Nigeria. These “jellies” are macroscopic, blue-green algal masses whose excessive mucilage of high consistency acts as substrata for other epiphytic components viz. diatoms and green algae. They are more abundant in the lentic areas of the lake, undergo diurnal vertical movements and have their growth enhanced by spikes of nitrogen and phosphorus salts into their aquatic medium. The effects of these “jellies” on the ecology and general water use of the lake are discussed. Speculations on the reasons for their occurrence are given and possible control measures outlined.     

Molecular study of two distinct phytoplasma species associated with streak yellows of date palm in Iran

A disease with symptoms similar to palm lethal yellowing was noticed in the early 2013 in Khuzestan Province (Iran) in date palm (Phoenix dactylifera). Infected trees displaying symptoms of streak yellows and varied in the incidence and severity of yellowing. A study was initiated to determine whether phytoplasma was the causal agent. Polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) methods using universal phytoplasma primers pairs R16mF1/mR1 and M1/M2 were employed to detect putative phytoplasma(s) associated with date palm trees. Nested PCR using universal primers revealed that 40 out of 53 trees were positive for phytoplasma while asymptomatic date palms from another location (controls) tested negative. RFLP analyses and DNA sequencing of 16S rDNA indicated that the presence of two different phytoplasmas most closely related to clover proliferation (CP) phytoplasma (group 16SrVI) and ash yellows (AY) phytoplasma (group 16SrVII). Sequence analysis confirmed that palm streak yellows phytoplasmas in each group were uniform and to be phylogenetically closest to “CandidatusP. fraxini” (MF374755) and “Ca. P. trifolii” isolate Rus‐CP361Fc1 (KX773529). Result of RFLP analysis of secA gene of positive samples using TruI and TaqI endonuclease is in agreement with rDNA analysis. On this basis, both strains were classified as members of subgroups 16SrVI‐A and 16SrVII‐A. This is the first report of a phytoplasma related to CP and AY phytoplasma causing date palm yellows disease symptoms.     

Molecular study of phytoplasmas associated with pistachio yellows in Iran

A large‐scale survey was conducted on pistachio plants exhibiting foliar symptoms including scorch, little leaf, yellows and reddish in pistachio growing areas in the Qom, Yazd and Qazvin provinces of Iran. Total DNA was extracted from symptomatic and symptomless pistachio and used in nested PCR assays with phytoplasma universal primers. Nested PCR products were obtained for symptomatic plant samples while the symptomless plants yielded no PCR products. Virtual restriction fragment length polymorphism, phylogenetic and DNA homology analyses of partial 16S ribosomal sequences of phytoplasma strains associated with symptomatic plants revealed the presence of phytoplasmas referable to two ribosomal groups; in particular, “Candidatus Phytoplasma solani” and “Ca. P. phoenicium” were identified. The presence of these phytoplasmas in pistachio is of great phytosanitary significance due to its commercial interest.     

Biophysical Examinations of the Bending Stability of Various Stele Types and the Upright Axes of Early “Vascular” Land Plants

Biophysical methods allow quantitative deductions as to the “bending effectivity” of various stele types and the contribution of different tissues to the stability of upright plant axes. Considering the first occurrence of the different stele types in geological history, a strong tendency towards a higher “bending effectivity” can be observed. It can be shown that Rhynia gwynne-vaughanii, Aglaophyton major (Rhynia major) and Asteroxylon mackiei as well as probably the vast majority of early “vascular” land plants were turgor systems, i.e. that the parenchyma (when fully turgescent) is by far the most important contributory factor towards the bending stability of the upright axes.     

Antagonistic effect of enterocin CCM 4231 from Enterococcus faecium on “bryndza”, a traditional Slovak dairy product from sheep milk

“Bryndza” is a traditional Slovak dairy product (type of soft cheese) made from sheep cheese which was ripened for 14 days. Because its manufacture, transporting and/or storing represent conditions which facilitate contamination, the effect of enterocin CCM4231 in “bryndza” was investigated with the aim to reduce the contaminant agents. “Bryndza” was divided into equal portions (50 g). The experimental sample (ES) as well as the control sample one (C1) were inoculated with Listeria innocua Li1 strain. The other control samples C2 and C3 were without Li1 strain. C3 control was selected as a reference control. ES and C2 portions were treated with purified enterocin CCM4231 in a concentration of 6400 AU/ml. Before the experimental inoculation, “bryndza” was checked for the presence of contaminant agents. The experiment lasted 1 week and the samples were stored in the refrigerator at 4 °C. Sampling was performed on day 1, on day 4 and on day 7. The control samples C2 and C3 were checked only on day 1 and then after 1 week. The following contaminant agents were detected in “bryndza” before its experimental inoculation with L. innocua Li1 strain: Escherichia coli in the amount 10³ cfu/ml/g, Staphylococcus aureus (10² cfu/ml/g) and enterococci (10⁴ cfu/ml/g). In the control sample C2, the number of E. coli was reduced to 10² cfu/ml/g. Enterococci and staphylococci were totally eliminated there. Concerning C3 control, natural decrease of bacteria was found and/or their unchanged counts. The value of pH (5) was stable during the whole experiment. In the experimental sample inoculated with Li1 strain, its counts were decreased immediately after enterocin CCM4231 addition approximately by one order of magnitude. This inhibitory effect was also detectable on day 4 by the difference of one order of magnitude between ES and C1. On day 7, 10³ cfu/ml/g of Li1 strain were detected in both samples (ES, C1). The difference by one order of magnitude indicated, an inhibitory effect of enterocin CCM4231 in “bryndza”. However, bacteriocin activity was not determined by laboratory analyses.     

Molecular systematics,species limits,and diversification of the genus Dendrocolaptes (Aves: Furnariidae): Insights on biotic exchanges between dry and humid forest types in the Neotropics

The true diversity and interspecific limits in the Neotropical endemic avian genus Dendrocolaptes (Furnariidae) remain a highly controversial subject, with previous genus‐wide assessments, based mostly on morphological characters, producing poorly resolved phylogenies. The lack of well‐resolved, robust, and taxonomically densely sampled phylogenies for Dendrocolaptes prevents reliable inferences on the genus’ actual species diversity and evolutionary history. Here, we analyzed 2,741 base pairs of mitochondrial and nuclear genes from 43 specimens belonging to all species and the majority of subspecies described for Dendrocolaptes to evaluate species limits and reconstruct its diversification through time. Our phylogenies recovered a monophyletic Dendrocolaptes, with two main highly supported internal clades corresponding to the D. certhia and D. picumnus species complexes. Also, our analyses supported the monophyly of most Dendrocolaptes species recognized today, except D. picumnus, which was consistently recovered as paraphyletic with respect to D. hoffmannsi. A coalescent‐based test supported a total of 15 different lineages in Dendrocolaptes and indicated that the number of currently accepted species within the genus may be greatly underestimated. Particularly relevant, when combined with previous analyses based on plumage characters, comparative high levels of genetic differentiation and coalescent analyses support the recognition of D. picumnus transfasciatus as a full species that is already under threat. Ancestral area reconstructions suggest that diversification in Dendrocolaptes was centered in lowland Amazonia, with several independent dispersal events leading to differentiation into different adjacent dry and high elevation forest types throughout the Neotropics, mainly during the Middle and Late Pleistocene.     

Reproductive patterns in galagos (Galago zanzibaricus and Galago garnettii) in relation to climatic variability

The reproductive patterns (birth seasonality, litter size, litters per year) of two sympatric species of galago (Galago zanzibaricus and G. garnettii) were studied in a coastal forest in Kenya for a two-year period. Trap-retrap and radio tracking methods were employed. G. zanzibaricus has one infant twice per year; G. garnettii has one infant once per year. Both species are seasonal breeders. These East African galagos are intermediate in reproductive patterns when compared with galagos from South African woodland (G. senegalensis moholi and G. crassicaudatus umbrosis) and West African rainforest (G. alleni and G. demidovii). Climatic patterns (total annual rainfall, seasonal variability of rainfall, variability in total annual rainfall, and annual temperature variability) are also compared for the three regions. Climatically, East Africa is intermediate between West and South Africa in total annual rainfall and in seasonality of rainfall, but not in year-to-year variability in rainfall. East Africa shows the highest variability in annual rainfall. South Africa has the coldest dry seasons and highest variability in temperatures. The results of this study suggest that “r-selection” and “K-selection” do not provide adequate explanations of galago reproductive patterns.     

Basic description and some notes on the evolution of seven sympatric morphs of Dolly Varden Salvelinus malma from the Lake Kronotskoe Basin

The study examines the basic morphological and ecological features of Dolly Varden from Lake Kronotskoe (Russia, Kamchatka). Seven valid morphs different in head proportions, feeding, timing, and place of spawning have been determined in this ecosystem. The basic morphometric characteristics clearly separate Lake Kronotskoe morphs from each other, as well as from its potential ancestor (Dolly Varden). According to CVA analysis, the most notable morphological characteristics determining the mouth position are the length of a lower jaw and rostrum. Furthermore, five of seven morphs inhabit different depth zones of the lake and feed on different food resources. Our data suggest that reproductive isolation may be maintained by temporal/spatial isolation for two morphs with lacustrine spawning, and by spatial isolation only for the rest of the morphs with riverine spawning. The sympatric diversity of the Lake Kronotskoe charrs is exceptionally wide, and there are no other examples for seven sympatric morphs of genus Salvelinus to coexist within a single ecosystem. This study puts forward a three‐step hypothetical model of charr divergence in Lake Kronotskoe as a potential ground for future studies.     

Growth of an Insect Trypanosomatid at 37 C in a Defined Medium*

SYNOPSIS. A defined medium for an insect trypanosomatid, “Leptomonas pessoai” (probably a member of the genus Herpetomonas) isolated from the reduviid Zelus leucogrammus, allows growth up to 37 C. No marked differences were evident for growth at 37 C. The requirements for amino acids, vitamins, purine, and hemin, and pH range were like those established for Crithidia fasciculata. Again like C. fasciculata, it remains alive at 4 C for at least 3 months in a çlycerolated defined medium.     

Early leaf miners and the ground plan of the lepidopteran larval trunk: Caterpillar morphology of the basal moths Heterobathmia,Eriocrania, and Acanthopteroctetes

The larval trunk morphology including chaetotaxy, locomotory structures, and trunk musculature of Heterobathmia pseuderiocrania, Eriocrania cicatricella, and Acanthopteroctetes unifascia is described using conventional light, polarization, and scanning electron microscopy. The ground plan morphology of the lepidopteran larva and neolepidopteran caterpillar is discussed in light of the life history succession from free soil dwelling organism to endophagous and finally to a primarily free living, angiosperm associated organism. I suggest that the larval morphology is argued to be strongly influenced by the shift in number of surfaces present in the larval environment. Especially the environment of the endophagous species, where the upper surface of the leaf mine is linked to the presence of dorsal locomotory structures such as the retractable calli and dorsal friction patches is proposed to have had a significant impact on the morphology and locomotory mechnism of the lepidopteran caterpillar. The chaetotaxy of the lepidopteran ground plan is found to be simple, consisting only of primary and secondary tactile setae and segmental proprioceptors. The presumption of Gerasimov ([1935] Zool Anz 112:177–194) that MXD1 of the prothorax is a shifted mesothoracic MD setae is supported. I suggest that the serial arrangement of the proprioceptors MD1, present on all trunk segments except the prothorax, and a trisetous MV group on all the thoracic segments is part of the lepidopteran larval ground plan. The absence of apodeme structures associated with trunk musculature in the nonglossatans suggests that this is an autapomorphic character of the Lepidoptera and it is further found to have been influential in the evolution of the typical caterpillar trunk. The attachments of the thoracic muscles directly to the trunk integument, suggest that the apodemal structures ancestral to the Amphiesmenoptera have been reduced in the Lepidoptera. Within the non‐Neolepidoptera, the lifehistory shift may have resulted in reduction of the dorsal locomotory structures, such as calli. The abdominal musculature and structural similarities further suggest that the ventral calli are structural predecessors to the crotchet bearing proleg of the “typical caterpillar.” J. Morphol. 274:1239–1262, 2013. © 2013 Wiley Periodicals, Inc.     

Unrecognized and imperilled diversity in an endemic barb (Smiliogastrini,Enteromius) from the Fouta Djallon highlands

The Upper Guinean Forests of Guinea, Sierra Leone and Liberia contain high levels of freshwater biodiversity. The Guinean Range and associated Fouta Djallon highlands separate two biogeographical provinces in the region and served as a refugium during past climatic fluctuations. While many species of freshwater fishes are restricted to one biogeographical province or the other, some are reported to occur on both sides of the divide. Here, we examine the molecular and morphological diversity of an endemic small African barb, Enteromius foutensis, reported to occur in both provinces. This integrative analysis revealed unrecognized diversity and suggests recent, or ongoing, events that facilitated geodispersal and subsequent divergence in the region. The molecular analysis revealed three divergent and well‐supported populations within E. foutensis. Accounting for allometric shape variation allowed us to observe diagnostic morphological differences among the populations. Enteromius foutensis sensu stricto is restricted to the Little Scarcies drainage in Guinea and northern Sierra Leone. Our study revealed two candidate species distinct from E. foutensis. One is likely a narrow endemic restricted to a small area in the Konkouré River basin; the other candidate species inhabits the upper Senegal and Gambie River drainages. How these patterns of diversity compare with other freshwater species from the Fouta Djallon highlands and the conservation status of these candidate species are also discussed.     

Significance and Interspecific Variability of Accumulated Trace Metal Concentrations in Antarctic Benthic Crustaceans

Trace metals (Cd, Cu, Pb and Zn) were analysed in crustaceans collected on Polarstern cruises ANT XVI/2 (1999) and ANT XXI/2 (2003/04, BENDEX) to the Weddell Sea. Our study provides further evidence for the frequently reported “Cd anomaly” in polar crustaceans, with data ranging from 1.2 (Ceratoserolis trilobitoides) to 6.2 mg Cd kg^–1 DW (Notocrangon antarcticus) in 1999 and from 1.2 (Waldeckia obesa) to 20.3 mg Cd kg^–1 (Tryphosella murrayi) in 2003. Pb concentrations well below 1 mg kg^–1 in most of the samples analysed might serve as a regional or even global background value for comparison in biomonitoring studies. Increasing Cu concentrations from eggs of decapods (e.g., 5 vs. 51 mg kg^–1 in N. antarcticus) or juveniles in the brood pouch of an amphipod species to adult females indicate that the enzymatic requirements and haemocyanin component demand for Cu in early life‐history stages is probably not met without a distinct bioaccumulation of this essential element after hatching. Most interestingly, Cd also increases (<0.1 vs. 6.2 mg kg^–1 in N. antarcticus). This could be the consequence of efficient uptake mechanisms for Cu that cannot discriminate between this element and Cd. Cu and Zn concentrations in decapods of this study are largely within the range reported worldwide (40–90 mg Cu kg^–1 and 40–80 mg Zn kg^–1), indicating that these elements are regulated. The enormous heterogeneity of Cd and Zn in many amphipod species investigated (e.g., from 0.6 in Gnathiphimedia mandibularis to 34.4 mg Cd kg^–1 in Orchomenopsis acanthura and from 41 in Eusirus antarcticus to 1244 mg Zn kg^–1 in Iphimediella bransfieldi) supports the hypothesis of the “Cd anomaly” and suggests that there is probably no consistent metabolic demand for the essential element Zn in this taxonomic group. The heterogeneity of Cu in amphipods is less pronounced. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Dimensions of face in Asia in the perspective of geography and prehistory

Data from 24 craniofacial dimensions were compiled for samples representing all the human populations of the world. These were converted into C scores and used to construct Euclidean Distance dendrograms. The populations of the world are best depicted as 8 major regional clusters representing: Africa, Amerind, Asia-Mainland, Australo-Melanesia, Eskimo-Siberia, Europe, India, and Jomon-Pacific. The mainland Asian cluster divided into northern and southern components. The Neolithic in both areas is less clearly associated with the northern or southern components and may represent the form of their undifferentiated precursors. This needs to be tested by the addition of further specimens and an adequate sample from South China. Mongols are peripheral members of the Asia-Mainland cluster and should not be thought of as typifying the region as a whole. The use of the term "Mongoloid" is inappropriate in both theory and practice.     

Rapid detection of “Candidatus Phytoplasma mali” by recombinase polymerase amplification assays

Isothermal recombinase polymerase amplification (RPA) assays for the specific detection of “Candidatus Phytoplasma mali (Ca. P. mali),” the causal agent of apple proliferation, were developed. The assays amplify a fragment of the imp gene and amplimers were detected either by fluorescence in real‐time mode (TwistAmp^®exo assay) using a fluorophore‐labelled probe or by direct visualization employing a lateral flow device (TwistAmp^®nfo assay/Milenia^®HybriDetect). The RPA assays specifically amplified DNA from “Ca. P. mali” strains, and cross‐reactivity with other phytoplasmas or plant DNA was not observed. The limit of detection was determined with a cloned imp standard, and positive results were obtained down to 10 copies with both RPA assay formats. In comparison with a TaqMan real‐time PCR assay based on the same target gene, the RPA assays were equally sensitive, but results were obtained faster. Simplified nucleic acid extraction procedures from plant tissue with Tris‐ and CTAB‐based buffers revealed that crude Tris–DNA extracts were a suitable source for RPA tests while larger concentrations of CTAB were inhibitory. This is the first report of RPA‐based assays for the detection of “Ca. P. mali”. The assays are suitable for high‐throughput screening of plant material and point‐of‐care diagnostic and can be potentially combined with a simplified DNA extraction procedure.     

A novel mcl-PHA produced on coprah oil by Pseudomonas guezennei biovar. tikehau, isolated from a "kopara" mat of French Polynesia

Pseudomonas guezennei biovar. tikehau was isolated from a microbial mat on the atoll of Tikehau in French Polynesia, and is able to synthesize medium chain length poly-beta-hydroxyalkanaote copolymers when grown on coprah oil. A two-step cultivation process was used and the biosynthesis of PHAs was followed along 52h by regular culture sampling. The polyester was purified from freeze-dried cells and analysed by nuclear magnetic resonance (NMR), Fourier transform infra red (FTIR), and gas chromatography mass spectrometries. The copolyester produced by P. guezennei biovar. tikehau from coprah oil mainly consisted of saturated monomers, i.e. 3-hydroxyoctanoate (3HO) and 3-hydroxydecanoate (3HD), and the monomeric composition of the polyester did not change during the fermentation process. However, yield of PHAs production varied from 4% of the cellular dry weight (CDW) to 63% obtained after 36h. Scan electron microscopy was used to study the morphology and organization of PHAs granules within the cells and revealed the presence of several granules occupying almost the entire cell volume.     

Proximate Composition and Nutrient Elements in the “Unusual” Algal “Jellies” of Lake Oguta in Southern Nigeria

The proximate composition, nutrient and trace elements of the planktonic algal “jellies” of Lake Oguta in Southern Nigeria, are provided as guides to their utilization and control, using flood (rainy) and dry season samples. These “jellies” are an unusual, macroscopic planktonic algal community in the lake, made up mostly of blue-green algae (Microcystis, Anabaena, Chroococcus) with some green algae (e. g. Spirogyra) and pennate diatoms (e.g. Navicula) rather epiphytic on the mucous jellies of the blue-green algae. Water content was high (98.2% mean, range 97.5-99.5%). Ash, protein and fat had values of 41.7 (39.0-46.2%), 16.9 (15.6-19.4%), and 2.0 (1.7-2.1%) respectively. Crude fibre and carbohydrates were high (25.8% and 12.4%). The levels of phosphorus and nitrogen were rather high, so were the major cations whose order of dominance (Ca > K > Mg > Na) differed from that of the lake water. Macronutrients increased in their concentrations during the rainy flood season while the reverse was the case with the micronutrients. Diagenesis of the endogenic minerals rather than anthropogenic factors are thought to be the reason for the seasonal changes in the ionic concentrations. Possible uses of the algal jellies are discussed as well as their control options.