Nectar feeding by the Cape rock elephant-shrew Elephantulus edwardii (Macroscelidea) — A primarily insectivorous mammal pollinates the parasite Hyobanche atropurpurea (Orobanchaceae)

Cape rock elephant-shrews, Elephantulus edwardii (Macroscelidea) were found to be pollinators of Hyobanche atropurpurea (Orobanchaceae), a member of a holoparasitic plant genus, lacking up to now any detailed pollinator observations. The elephant-shrews were live-trapped near the plants in the Western Cape of South Africa. One animal was released in a terrarium, containing the plants. It licked nectar with its long and slender tongue while being dusted with pollen and touching the stigmas of the flowers with its nose and the fur above the snout. H. atropurpurea pollen was found in swabs of the fur around noses and snouts as well as in the faeces of captured animals. The occurrence of pollen in the faeces of the elephant-shrews is likely a result of grooming their fur as they visited the flowers without eating or damaging them. This is one of the first records of pollinating, nectar-drinking elephant-shrews, contributing to the very scarce knowledge about the behaviour of those primarily insectivorous animals, as well as small mammal pollination.     

Comparative phytochemistry of eleven species of Vigna (Fabaceae)

A survey of the biochemical constituents of 11 species of Vigna indicates the absence of the non-protein amino acid canavanine in their seeds, and absence of proanthocyanidin (polyphenol) in their leaves. Proanthocyanidin was found in the seeds of all, except Vigna subterranea. The constitutive leaf flavonoids of four genotypes of the pantropic V. subterranea were also studied and compared with those from three other cultivated species. The flavonoid kaempferol seems to be most prevalent as it was found in all of the four cultivated species and genotypes. The glycoside kaempferol-3-O-rutinoside was found present in the four genotypes of V. subterranea and other cultivated Vigna species. However, the flavonoid kaempferol-3-O-glucoside-7-rhamnoside is restricted to V. subterranea. This study questions the inclusion of V. subterranea in the genus Vigna on account of absence of seed proanthocyanidin and restricted accumulation of kaempferol-3-O-glucoside-7-rhamnoside in the leaves.     

A rare new species of Polhillia (Genisteae,Fabaceae)

The new species Polhillia ignota Boatwr. is described. This species is known only from two collections, one between Vredenburg and Saldanha and the other close to Porterville. The new species is most similar to P. obsoleta, which is known only from a few localities around Worcester, in its narrow, sericeous leaflets and flowers of equal size, but differs in its flattened mature leaves with larger leaflets (up to ± 12 mm long), longer pseudo-peduncles (up to ± 15  mm long), denser inflorescences (with up to four flowers), shorter pedicels (1–2 mm long) and non-auriculate wing- and keel petals.     

Hybridizations and genetic relationships among Lindernia species (Scrophulariaceae): L. procumbens and two subspecies of L. dubia

Lindernia procumbens and L. dubia are common annual weeds in flooded rice fields of Japan. Two subspecies of L. dubia, subsp. major and subsp. dubia, are usually recognized in Japan but they are both regarded as synonyms of L. dubia elsewhere. In a cluster analysis based on AFLP, most L. dubia subsp. major formed a separate cluster from L. dubia subsp. dubia although 11% of haplotypes classified using AFLP were not coincident with classification using the shape of leaf bases, which is the commonly used identification trait. Artificial F₁ plants between L. procumbens and L. dubia subsp. major, and between L. procumbens and L. dubia subsp. dubia did not produce seed. Forty percent of capsules produced by F₁ plants from these two subspecies were slimmer and 80% pollen were sterile in slimmer capsules. However, seed number of most F₁ capsules was not different from that of self-fertilized plants, suggesting that there was no complete reproductive isolation between the subspecies. Natural hybridization of these subspecies may have occurred but we are not aware of it because F₁ plants are rare and F₂ plants are indistinguishable from these subspecies.     

Reproductive biology and pollination of southeastern Brazilian Stanhopea Frost ex Hook. (Orchidaceae)

Reproductive biology and pollination of Stanhopea lietzei and Stanhopea insignis were studied in a semi-deciduous mesophytic forest in the Serra do Japi (SJ), and in the coastal plain of Picinguaba, both in the State of São Paulo, Brazil. Floral morphology, pollination, breeding system and fruit set of both species were investigated. S. lietzei and S. insignis are pollinator-specific, being pollinated by male bees of Eufriesea (Apidae, Euglossini), which collect the fragrance produced by pluricellular osmophores at the base of the saccate hypochile. S. lietzei and S. insignis were pollinated by Eufriesea pulchra and Eufriesea purpurata, respectively. Observations using substances present in the floral fragrance of both studied species as chemical baits were also performed. E. purpurata was attracted by benzyl alcohol, the major compound of the perfume of S. insignis, while E. pulchra was attracted by none of the compounds used. Both studied Stanhopea are self-compatible but pollinator dependent. Self-pollination, however, tends to be avoided by floral mechanisms. In experimental self- and cross-pollinations the proportion of fruit abortion was high and related to resource limitation. The reproductive success of S. lietzei and S. insignis was low as a consequence of deficient pollen transference while pollinator scarcity was the main factor.     

Mycoplasma feriruminatoris sp. nov., a fast growing Mycoplasma species isolated from wild Caprinae

Five Mycoplasma strains from wild Caprinae were analyzed: four from Alpine ibex (Capra ibex) which died at the Berlin Zoo between 1993 and 1994, one from a Rocky Mountain goat collected in the USA prior to 1987. These five strains represented a population different from the populations belonging to the ‘Mycoplasma mycoides cluster’ as tested using multi locus sequence typing, Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and DNA–DNA hybridization. Analysis of the 16S rRNA gene (rrs), genomic sequence based in silico as well as laboratory DNA–DNA hybridization, and the analysis of phenotypic traits in particular their exceptionally rapid growth all confirmed that they do not belong to any Mycoplasma species described to date. We therefore suggest these strains represent a novel species, for which we propose the name Mycoplasma feriruminatoris sp. nov. The type strain is G5847^T (= DSM 26019^T = NCTC 1362^T).     

Pollen tube reallocation in two preanthesis cleistogamous species, Ranalisma rostratum and Sagittaria guyanensis ssp. lappula (Alismataceae)

Pollen deposition on stigmas and pollen tube growth in two apocarpous species, Ranalisma rostratum and Sagittaria guyanensis ssp. lappula (Alismataceae), were examined with fluorescence microscopy. The reallocation of pollen tubes among pistils was observed in both species. The percentage of pollinated stigmas per flower was only 22.0% in R. rostratum and 51.0% in S. guyanensis, though the seed/ovule ratios are higher than 65% in both species. The number of pollen grains on each single stigma ranged from 0 to 96 in R. rostratum, and from 0 to 125 in S. guyanensis. When more than one pollen grain deposited on a stigma, all pollen tubes grew to the ovary, but only one of them turned towards the ovule and finally entered the nucleus. The other tubes grew through the receptacle tissue into ovules of adjacent carpels whose stigmas were unpollinated or pollinated later. The intercarpellary growth of pollen tubes could be a mechanism to increase the efficiency of sexual reproduction in an apocarpous gynoecium with low pollination on the pistils.     

Isoenzyme variation patterns and species concept in Astragalus gossypinus and Astragalus persicus complexes (Fabaceae) in Iran

Isoenzyme electrophoresis was employed to examine the relationships of 21 individuals representing four populations of Astragalus gossypinus complex as well as 20 individuals representing five populations of Astragalus persicus complex. A total of 27 bands from three enzyme systems (superoxide dismutase, esterase, and peroxidase) were obtained. UPGMA clustering method resulted in two distinct clusters for different populations corresponding to the two species complexes analysed. In both species, populations distributed on Alborz mountain range in northern Iran form separated clusters from those distributed on Zagros mountain range in the West. The results also show that both species complexes exhibit a high diversity based on Shannon–Weaver diversity index (H′) compared with other species of Astragalus reported previously. The mean number of bands per each presumed isoenzyme ranges from 2.14 to 3.57. The value of Euclidean distance ranges from 1.251 to 3.152. Our data suggest that both species should be circumscribed wider than that treated by most taxonomists, and several taxonomic names should be reduced under synonymy of the corresponding species.     

Pollination biology of Indotristicha ramosissima (Podostemaceae: Tristichoideae)

Floral morphology, phenology and mode of pollination have been studied in Indotristicha ramosissima (Wt.) van Royen. Although the plants are submerged, self-pollination (autogamy) occurs above water. This is aided by considerable elongation of the pedicel (20 mm) prior to pollination. The filaments of the stamens also elongate rapidly (∼6.5 mm/h) before and after pollination. The flowers are typically trimerous. Each anther contains 1743 ± 187 pollen grains. These are spherical, multiporate, 3-celled and ∼97% fertile at the time of shedding. Germination of pollen on the stigma and the growth of pollen tubes have been traced in both naturally and manually (self- as well as cross-) pollinated pistils. The pollen:ovule ratio is ∼72:1 and the ovule: seed ratio is ∼2:1. Mature fruits are 8 or 9 ribbed and open by 2 or 3 longitudinal slits that release ∼32 seeds.     

Molecular and morphological relationships between two closely related species, Turbinaria ornata and T. conoides (Sargassaceae,Phaeophyceae)

We employed four markers (two chloroplastic, one mitochondrial and one nuclear) to investigate whether Turbinaria ornata and Turbinaria conoides, two morphologically similar brown algae, are distinct species. The molecular data revealed reciprocal monophyly, thus confirming the two algae as discrete species. Nevertheless, taxonomical ambiguities remain. T. conoides is more morphologically variable than T. ornata. We identified an atypical T. conoides morphotype whose blade possesses intramarginal teeth, a morphological characteristic of the T. ornata group that is normally absent within T. conoides. The molecular data indicate, nonetheless, that this morphotype belongs genetically to the T. conoides group. Thus, the results cast doubt upon the validity of using intramarginal teeth for species diagnosis.     

RPB2 sequences reveal a close phylogenetic relationship between tetraploid Hordelymus and diploid Hordeum species in Triticeae (Poaceae)

The origin of Hordelymus genome has been debated for years, and no consensus conclusion was reached. In this study, we sequenced and analyzed the RPB2 (RNA polymerase subunit II) gene from Hordelymus europaeus (L.) Harz, and its potential diploid ancestor species those were suggested in previous studies. The focus of this study was to examine the phylogenetic relationship of Hordelymus genomes with its potential donor Hordeum, Psathyrostachys, and Taeniatherum species. Two distinguishable copies of sequences were obtained from H. europaeus. The obvious difference between the two copies of sequences is a 24 bp indel (insertion/deletion). Phylogenetic analysis showed a strong affinity between Hordeum genome and Hordelymus with 85% bootstrap support. These results suggested that one genome in tetraploid H. europaeus closely related to the genome in Hordeum species. Another genome in H. europaeus is sister to the genomes in Triticeae species examined here, which corresponds well with the recently published EF-G data. No obvious relationship was found between Hordelymus and either Ta genome donor, Taeniatherum caput-medusae or Ns genome donor, Psathyrostachys juncea. Our data does not support the presence of Ta and Ns genome in H. europaeus, and further confirms that H. europaeus is allopolyploid.     

Hydrozoan and protozoan epibionts on two decapod species, Liocarcinus depurator (Linnaeus, 1758) and Pilumnus hirtellus (Linnaeus, 1761), from Scotland

Crabs of the species Liocarcinus depurator and Pilumnus hirtellus, collected on the west coast of Scotland, showed hydrozoan and protozoan epibionts. The epibionts of Liocarcinus depurator were the protozoan Ephelota plana and the hydrozoans Clytia gracilis and Leuckartiara sp. The epibionts of Pilumnus hirtellus were the protozoans E. plana and Zoothamnium sp. For Liocarcinus depurator the number of Ephelota per crab fluctuated between 0 and 47 and the greatest number of suctorians were located on the chelipeds, carapace and anterior pereiopods. The hydrozoans, for Liocarcinus depurator, showed densities of 0-20 (Clytia gracilis on the second pereiopod) and 0-507 individuals per crab (Leuckartiara sp., principally on chelipeds, carapace and the fourth right pereiopod). For Pilumnus hirtellus, Ephelota plana showed densities between 3 and 56 individuals per crab, the greatest number of suctorians being located on the same areas as on Liocarcinus depurator. There was a density of 10-69 individuals per crab of Zoothamnium sp. on Pilumnus hirtellus (located on the carapace). Ephelota plana has not been observed previously as an epibiont on crustacea, nor had Zoothamnium sp. as an epibiont on Pilumnus hirtellus. Both hydrozoans, Leukartiara and Clytia, have not been previously described as epibionts on Liocarcinus depurator. Data about the morphological characteristics and distribution of these epibionts are included.     

Comparative cryopreservation study of trochophore larvae from two species of bivalves: Pacific oyster (Crassostrea gigas) and Blue mussel (Mytilus galloprovincialis)

Oysters and mussels are among the most farmed species in aquaculture industry around the world. The aim of this study was to test the toxicity of cryoprotective agents to trochophore larvae from two different species of bivalves and develop an improved cryopreservation protocol to ensure greater efficiency in the development of cryopreserved trochophores (14 h old oyster larvae and 20 h old mussel larvae) to normal D-larvae for future developments of hatchery spat production. The cryopreservation protocol producing the best results for oyster trochophores (60.0 ± 6.7% normal D-larvae) was obtained by holding at 0 °C for 5 min then cooling at 1 °C min⁻¹ to −10 °C and holding for 5 min before cooling at 0.5 °C to −35 °C, holding 5 min and then plunging into liquid nitrogen (LN), using 10% ethylene glycol. For mussel experiments, no significant differences were found when cooling at 0.5 °C min⁻¹ or at 1 °C min⁻¹ for CPA combinations with 10% ethylene glycol and at 0.5 °C min⁻¹. Using these combinations, around half of trochophores were able to develop to normal D-larvae post-thawing (48.9 ± 7.6% normal D-larvae).     

Centrosema is a promiscuous legume nodulated by several new putative species and symbiovars of Bradyrhizobium in various American countries

Centrosema is an American indigenous legume that can be used in agroecosystems for recovery of acidic and degraded soils. In this study, a Centrosema-nodulating rhizobial collection of strains isolated in a poor acid savanna soil from Venezuela was characterized, and the members of the collection were compared to other Centrosema strains from America. The analysis of the rrs gene showed that the strains nodulating Centrosema in American countries were closely related to different species of the genus Bradyrhizobium. However, the analysis of the atpD and recA genes, as well as the 16S–23S ITS region, showed that they formed several new phylogenetic lineages within this genus. The Venezuela strains formed three lineages that were divergent among themselves and with respect to those formed by Centrosema strains isolated in other countries, as well as to the currently described species and genospecies of Bradyrhizobium. In addition, the symbiotic genes nodC and nifH carried by Centrosema-nodulating strains were analyzed for the first time, and it was shown that they belonged to three new phylogenetic lineages within Bradyrhizobium. The nodC genes of the Centrosema strains were divergent among themselves and with respect to the genistearum and glycinearum symbiovars, indicating that Centrosema is a promiscuous legume. According to these results, the currently known Centrosema-nodulating strains represent several new putative species and symbiovars of the genus Bradyrhizobium.     

Comparative pollen morphology of Brachylena, Tarchananthus and two species of Tubulifloridites (Asteraceae) from the Eocene, Knysna Lignite of South Africa

Two fossil taxa Tubulifloridites antipodica and T. viteauensis recovered from the Eocene Knysna Lignite of South Africa were examined with scanning (SEM) and transmission electron microscopy (TEM). The details of their sculpturing and wall structure are similar to the same species of fossil dispersed pollen taxa recovered from southwestern Africa and South America. Fifteen species of the woody South African taxa, Brachylaena (9 species) and Tarchonanthus (6 species) were investigated with SEM and TEM. All of the taxa are tricolporate, spherical to slightly prolate, microechinate to echinate and have a bilayered columellate infrastructure, except B.ilicifolia, which has a single columellate infrastructural level with the “granularization” of the outer portion of the infrastructural layer or the inner layer of the tectum. There is a similar distribution of plesiomorphic and derived pollen characters in a number of aster subfamilies and tribes suggesting a similar evolutionary progression of pollen, and pollen wall character evolution was occurring synchronously in a variety of aster subfamilies during the middle Tertiary and that these unique pollen features may be important to the evolution and diversification of the Asteraceae.     

Essential oil composition of Achyrocline flaccida (Weinm.) DC. (Asteraceae) from different locations of Argentina

The qualitative and quantitative composition of the essential oils obtained from the inflorescences of Achyrocline flaccida (Asteraceae) has been investigated for the first time. Plant material was collected from eleven locations in Argentina. The essential oils were obtained by hydrodistillation (0.1–0.8% v/w, dried material) and analyzed by GC–FID–MS. Eighty-three compounds were identified representing more than the 90% of the oils. The major components were α-pinene and β-caryophyllene. Statistical analysis was performed in order to evaluate the variability of the essential oils analyzed. Two groups were formed reflecting only quantitative differences in the content of major compounds. The chemical pattern of essential oils observed for A. flaccida is similar to other Achyrocline species studied, except Achyrocline hyperchlora.     

Fruit anatomy of species of Solanum sect. Acanthophora (Solanaceae)

The fruits of 10 species of Solanum sect. Acanthophora were studied. Cross and/or longitudinal and/or tangential microtome sections, stained mostly with astra blue/basic fuchsin, were made for microscopic examination. Three different kinds of cells were found in the epidermis, immediately below which a hypodermis, consisting in any of four types of structures, was always found. The mesocarp exhibits two histologically differentiated zones, an external one (formed by normal or spongy parenchyma, according to the species), and an internal one, commonly juicy, and with proliferations among the seeds. The diagnostic value of all these structures is assessed. Morpho-anatomical information is used to define fruit types beyond the berry, traditionally described for Solanum, and the probable dispersal syndrome related to them is discussed. Fruit similarities are slightly noticeable in several cases, while differences may be the result of their sexual system – which affects specially the size – and their histology, which is related to the dispersal syndrome. The comparison of our data with previous molecular phylogeny of the section suggests that a significant morphological variation is not associated with significant DNA sequence changes.     

Giardia/giardiasis — A perspective on diagnostic and analytical tools

Giardiasis is a gastrointestinal disease of humans and other animals caused by species of parasitic protists of the genus Giardia. This disease is transmitted mainly via the faecal–oral route (e.g., in water or food) and is of socioeconomic importance worldwide. The accurate detection and genetic characterisation of the different species and population variants (usually referred to as assemblages and/or sub-assemblages) of Giardia are central to understanding their transmission patterns and host spectra. The present article provides a background on Giardia and giardiasis, and reviews some key techniques employed for the identification and genetic characterisation of Giardia in biological samples, the diagnosis of infection and the analysis of genetic variation within and among species of Giardia. Advances in molecular techniques provide a solid basis for investigating the systematics, population genetics, ecology and epidemiology of Giardia species and genotypes as well as the prevention and control of giardiasis.