Rodent pollination in the Cape legume Liparia parva

Flowers of Liparia parva from which rodents were excluded had lower seed‐set than open flowers. The rodent Acomys subspinosus was captured in the vicinity of this plant species and captures had substantial numbers of L. parva pollen in their scats. Captured individuals of A. subspinosus visited L. parva flowers in tanks and removed standard petals to obtain the nectar. Typical of rodent‐pollinated species, L. parva, flowered in winter and flowers mostly opened in the evenings and the stigma‐nectar distance was about 10 mm. This is the first evidence for rodent pollination in the large cosmopolitan family, the Fabaceae.     

Impact of Asymmetrical Hybridization Followed By Sterile F1 Hybrids on Species Replacement in Pseudorasbora

Pseudorasbora pumila, one of the endangered freshwater minnows in eastern Japan, has been largely replaced by the accidentally introduced species, P. parva, which originated from western Japan. In the contact zone, P. pumila and P. parva have hybridized intensively, producing sterile F₁ hybrids. The present study determined the maternal parent of F₁ hybrids using mtDNA haplotypes to investigate the mating system between P. parva and P. pumila in the hybrid zone. We also pursued the successive changes in the genetic structures of hybridizing populations over a 5-year period using allozymes. A total of 100 natural F₁ hybrids collected from six different populations had P. pumila mtDNA without exception, suggesting that sterile F₁ hybrids resulted from mating only between P. pumila females and P. parva males. Such asymmetrical hybridization implies that P. pumila females waste considerably greater reproductive efforts compared with P. parva males. The data suggest that the rapid replacement of P. pumila by P. parva has been promoted by asymmetrical hybridization, resulting in sterile F₁ hybrids.     

Genetic structures of allopatric and sympatric populations in Pseudorasbora pumila pumila and Pseudorasbora parva

Pseudorasbora pumila pumila was collected to investigate its genetic relationships from 11 localities in Hokkaido and eastern Honshu of Japan with Pseudorasbora parva from 6 localities in Hokkaido, eastern Honshu, and Polovynka River (a tributary of the Amur River in eastern Russia) in 1996–1998. In Hokkaido and Akita prefectures (Honshu), three P. pumila pumila populations were distributed with P. parva sympatrically. Samples of P. pumila pumila and P. parva were analyzed based on 27 allozyme loci. Genetic differentiation between P. pumila pumila and P. parva (D = 0.522–0.622, average 0.601) was almost equal to or greater than that between other congeneric fish species previously reported. In the three sympatric populations of two species, which had arisen from artificial transplantation of the latter into localities of the former, the individual frequency composition of genetic types (P. pumila pumila, P. parva, F₁ hybrid, and later filial generation hybrid) were significantly different from each other. Differing from the previous studies, F₁ hybrids were fertile and coexisting populations of the two species included the F₁ hybrid and the later filial generation hybrid. These results suggest that genetic population dynamics of sympatric populations of the two species would be influenced not only by hybridization but also by ecological and environmental factors. Alternatively, the existence of populations that possessed both the alleles specific to P. pumila pumila and those specific to P. parva may indicate that introgressive hybridization, instead of species displacement, has occurred between the two species under a certain condition.     

Towards the successful control of the invasive <Emphasis Type="Italic">Pseudorasbora parva</Emphasis> in the UK

The aim of this paper is to detail a control programme of the invasive Asian cyprinid fish Pseudorasbora parva in the UK that was initiated in March 2005. Described as Europe’s most invasive fish, P. parva presents a risk to native fishes through the transmission of a novel pathogen and undesirable impacts arising from processes including increased inter-specific competition. Populations have been recorded in 32 UK waters since their first recording outside of aquaculture in 1996; the majority are lakes <5 ha used for recreational angling in England. The aims of the control programme were to develop a basic evaluation framework that assesses populations by risk (high, medium and low), determine commensurate management actions according to that risk and then execute those actions. For populations assessed as ‘high-risk’, for example those that could result in P. parva dispersal into a river catchment, eradication was determined as the commensurate management action and six operations have since been completed, principally using rotenone, and all have been successful to date. For P. parva populations in sites evaluated as lower risk, techniques such as biomanipulation were determined as more appropriate actions and have been used to successfully reduce their abundance by >99%. To date, the total direct cost of this programme of sustained and on-going P. parva control is approximately ￡190,000.     

Loop-mediated isothermal amplification (LAMP) assays for detection of Theileria parva infections targeting the PIM and p150 genes

We have developed two loop-mediated isothermal amplification (LAMP) assays for the detection of Theileria parva, the causative agent of East Coast fever (ECF), an economically important cattle disease in eastern, central and southern Africa. These assays target the polymorphic immunodominant molecule (PIM) and p150 LAMP genes. The primer set for each gene target consists of six primers, and each set recognises eight distinct regions on the target gene to give highly specific detection of T. parva. The detection limit of each primer set is 1 fg, which is equivalent to one copy of the PIM and p150 T. parva genes. These PIM and p150 LAMP primer sets amplify DNA of T. parva isolates from cattle and buffalo from different countries including Kenya, South Africa, Tanzania, Rwanda, Uganda and Burundi, indicating their ability to detect T. parva from different countries. With the advantages of simplicity, rapidity and cost effectiveness, these LAMP assays are good candidates for molecular epidemiology studies and for monitoring control programs in ECF-endemic, resource poor countries.     

Size-dependent male–male competition for a spawning substrate between <Emphasis Type="Italic">Pseudorasbora parva</Emphasis> and <Emphasis Type="Italic">Pseudorasbora pumila</Emphasis>

Pseudorasbora parva, a species native to western Japan, has been accidentally introduced into eastern Japan, where P. pumila is indigenous. We investigated inter- and intraspecific, male–male competition between P. parva and P. pumila for acquisition of spawning substrates in an experimental setting. Within each species, males of larger standard length and heavier body weight were more successful in acquiring a substrate. Males of the two species competed, but the outcome was determined primarily by body weight. This interspecific, size-dependent, male–male competition might be an important factor in the species replacement of P. pumila by P. parva.     

Identifying conservation priority ponds of an endangered minnow, <Emphasis Type="Italic">Pseudorasbora pumila</Emphasis>, in the area invaded by <Emphasis Type="Italic">Pseudorasbora parva</Emphasis>

We investigated the occurrence pattern of the pond-living endangered cyprinid, Pseudorasbora pumila, and also compared its habitat characteristics with those of the congeneric invasive species, Pseudorasbora parva, in the contact zone. Comparison of 16 environmental variables among the P. pumila habitats, P. parva habitats, and unoccupied ponds revealed that conductivity was a common limitation factor of distribution of both species. We found that emergent vegetation occupancy along the pond bank was the most important factor determining P. pumila occurrence and that ponds with steep banks may have a low probability of containing P. parva. We constructed a logistic regression model to predict the establishment risk of P. parva in ponds occupied by P. pumila. The model demonstrated that more than half of the ponds exhibited a high establishment risk of P. parva. Principal component analysis using six parameters selected from stepwise logistic regression analysis revealed that seven unoccupied ponds had the potential to sustain P. pumila, suggesting that our study site is capable of supporting more P. pumila populations and expanding the current range.     

Isolation and characterization of polymorphic microsatellite DNA markers in topmouth gudgeon,Pseudorasbora (Teleostei: Cyprinidae)

Due to accidental introductions, the distribution of the east Asian topmouth gudgeon, Pseudorasbora parva, has rapidly expanded over recent years, thereby threatening the endangered Japanese species, P. pumila pumila. Sixteen microsatellite loci were isolated and characterized for P. parva, 14 being highly polymorphic (mean H_E = 0.58, mean number of alleles = 4.4). Successful characterization of 10 of these loci was also achieved for P. pumila pumila. These markers should be useful in future investigation of colonization of P. parva and the development of conservation strategies for P. pumila pumila.     

Sensitivity of the surface coat of the halotolerant green alga Dunaliella parva (Volvocales, Chlorophyceae) to lysozyme

The surface coat of Dunaliella parva Lerche was investigated using several techniques. Degradation by several cell lytic enzymes and ultrastructural observation revealed that D. parva has a specialized cell surface structure containing a glycoprotein that is sensitive not only to proteinases but also to lysozyme. This sensitivity was also demonstrated by electrophoresis of the cells and measurement of released glycerol after enzyme treatment. Immunochemical labeling indicated that the surface glycoprotein of D. parva is analogous to pepti-doglycan.     

A TEST FOR ENVIRONMENTAL EFFECTS ON BEHAVIORAL ISOLATION IN TWO SPECIES OF KILLIFISH

Behavioral isolation is a common and potent mechanism of reproductive isolation. Determining the extent to which behavioral isolation varies with environmental conditions is critical to understanding speciation and the maintenance of species boundaries. Here, we tested the effect of salinity on behavioral isolation (female species recognition, male–male competition, and male species recognition) between two closely related killifish (Lucania goodei and L. parva) that differ in salinity tolerance. We performed no‐choice assays and behavioral trials where males could compete and court females in fresh water (0 ppt) and brackish water (15 ppt). We found high levels of behavioral isolation that did not vary as a function of salinity. In behavioral trials, male species recognition of females was strong and asymmetric between the two species. Lucania goodei males preferred conspecifics and rarely courted or mated with L. parva females. Lucania parva males preferred conspecifics but readily courted and mated with L. goodei females. This asymmetry matches previously documented asymmetries in hybrid offspring fitness. Crosses between L. parva males and L. goodei females produce fully viable/fertile hybrids, but crosses between L. goodei males and L. parva females produce males with reduced fertility. Hence, behavioral isolation may have evolved in part due to reinforcement.     

Isolation of disproportionating enzyme from halotolerant microalga Dunaliella parva (Volvocales, Chlorophyceae)

Starch metabolism in Dunaliella parva Lerche is regulated by the osmolarity of the surrounding solute. Two isozymes showing amylolytic activity were obtained after purification by gel filtration chromatography. The isozymes show disproportionating activity (D‐enzyme) that is specific for malto‐oligosaccharides as substrate. Properties of the D‐enzyme in D. parva are similar to those in higher plants. The activity of the D‐enzyme is also found in various Dunaliella and Chlamydomonas, indicating that the D‐enzyme is also important in the starch metabolism in algae.     

Dunaliella acidophila: an algae with a positive zeta potential at its optimal pH for growth*

Abstract. The zeta potential (which approximates the surface potential) of the acid resistant green alga Dunaliella acidophila (optimal growth at pH 1.0) and the salt resistant D. parva (grown at pH 7.6) were calculated from the electrophoretic mobility of cells as determined by means of free-flow electrophoresis. Dunaliella acidophila cells exhibit a positive zeta potential (+5 to +20mV) at acidic external pH values, whereas negative zeta potentials (-30 mV) were measured at neutral pH values. Negative zeta potentials of the same order of magnitude were also measured for D. parva cells (pH 7.6). Low concentrations of La³⁺ and A1³⁺ did not affect the positive zeta potential of D. acidophila at acidic pH values, whereas higher concentrations caused a shift to more positive potentials. However, at neutral pH these cations caused a significant decrease of the negative zeta potential. The impermeant polycation poly-L-lysine acted in a similar manner to A1³⁺ or La³⁺. The effect of Impermeant cations and anions on various physiological reactions also supports the existence of a positive zeta potential for D. acidophila and of a negative zeta potential for D. parva: polycations such as DEAE-dextran and poly-L-lysine strongly inhibitied photosynthesis and mobility of D. parva, but did not affect these reactions in D. acidophila. Comparable differential inhibitions were also observed for A1³⁺ and La³⁺. Impermeant anions such as Dextran-sulfate exhibited effects in the opposite direction: inhibition was stronger with D. acidophila and weaker with D. parva. However, the differential inhibition by impermeant anions was much less pronounced in comparison with impermeant cations due to the relatively high pK_a values of anionic solutes and consequently relatively high protonation at pH 1.0. The physiological consequences of an asymmetrically charged plasma membrane (excess of positive charges outside, excess of negative charges on the cytoplasmic side) of D. acidophila are discussed in regard to the extreme acid resistance of this alga and its resistance to cationic toxic solutes in industrial wastes.     

Publications of Anthony Calhoun Clement (1909–1984)

Summary

Two zoeae and the megalopa of the majid crab Macropodia parva were obtained from laboratory material. At 25±1°C and 35% salinity, the first crab appeared 12 days after hatching; survival to the first crab instar was 27%. The larval stages are described in detail and compared with those of the previously described species of the genus Macropodia. Zoeal characteristics of M. parva that differentiate it from other known Macropodia larvae are the naked telson furcae and the absence, in zoea II, of the exopodal setae on the basis of the maxillule.     

Potential of Allolobophora parva (Oligochaeta) in vermicomposting

The aim of this study was to assess the potential of Allolobophora parva Eisen as a candidate for vermicomposting practices. Five organic waste mixtures: cow dung (CD), biogas plant slurry (BGS), cow dung + vegetable waste (CD + VW), BGS + VW and VW + Soil were vermicomposted using A. parva. Vermicomposting showed a decrease in pH, organic C and C:N ratio, but increase total N, available P and exchangeable K at the end. C:N ratio of end material (vermicompost) was within the agronomic acceptable limit (<20). The high level of NPK in worm-processed material indicates the candidature of this species for waste management operations. The earthworm also showed an excellent growth in different wastes. Results thus indicate that A. parva appeared a potential tool for conversion of organic wastes into value added products for sustainable land restoration practices.     

Survival of Theileria parva-infected adult Rhipicephalus appendiculatus under laboratory and quasi-natural conditions

Adult Rhipicephalus appendiculatus Muguga, having high or low intensities of Theileria parva Muguga infection in their salivary glands, were exposed to 20 °C and 85% relative humidity in the laboratory or quasi-natural conditions. Survival of the ticks and T. parva infections in their salivary glands was then monitored over a two year period. Ticks, having an average infection level of 2 infected acini per female, survived for up to 70 or 106 weeks after moulting under the laboratory or quasi-natural conditions respectively. Those having an infection level of 26 infected acini per female, survived for a similar duration except that those under quasi-natural conditions survived for a slightly shorter duration (102 weeks). Similarly, T. parva parasites survived for much longer periods under quasi-natural conditions than under the laboratory conditions. They survived for up to 38 or 78 weeks post salivary gland infection under the laboratory or quasi-natural conditions respectively in both categories of infection levels. There was apparently a density dependent relationship in T. parva survival, with a dramatic fall in infection occurring in ticks with high levels of infection between weeks 10 and 18 or weeks 38 and 46 post salivary gland infection in those exposed to laboratory or quasi-natural conditions before levelling off. This revised version was published online in July 2006 with corrections to the Cover Date.     

Overlapping niches between two co-occurring invasive fish: the topmouth gudgeon Pseudorasbora parva and the common bleak Alburnus alburnus

Invasive fish species impact aquatic ecosystems and modify native communities, often leading to a decline in local species. These ecological impacts include the transmission of pathogens, predation, competition as well as hybridization. Two invasive fish species, the common bleak Alburnus alburnus and the topmouth gudgeon Pseudorabora parva, have both been recently found co-occurring in several regions of southern Europe, such as the Italian Arno River. Nonetheless, the trophic relationships among invasive fish species, especially cyprinids, remain poorly understood, and no studies have reported the trophic interaction between these two species. This study compared length–weight relationship and used stomach content and stable isotope analysis of two co-occurring populations in the Arno River to characterize the growth and overlap of potential trophic niches. It also found similar allometric growth in both species, a wider generalist trophic niche for P. parva and a more specialized niche for A. alburnus. A considerable niche overlap was found, suggesting that feeding competition can occur if resources were to be limited. Moreover, the niche of P. parva was more likely to overlap with that of A. alburnus than vice versa, suggesting that P. parva can be considered as a potential over competitor. Nonetheless, the authors found in the overlapping populations no evidence of realized competition, probably avoided through a combination of fine-scale mechanisms. They also highlighted that these two invasive species can co-exist and share resources, at least in an open ecosystem like a river, thus potentially doubling up their trophic impact on local communities.     

The effects of sodium chloride,potassium chloride and glycerol on the activity of nitrate reductase of a salt-tolerant and two non-tolerant plants

Summary The activity of nitrate reductase from the salt-tolerant alga Dunaliella parva is inhibited by sodium chloride and potassium chloride, but not by glycerol. The activity of the enzyme from Chlorella pyrenoidosa Chick 611-8b and from the XD line of tobacco (Nicotiana tabacum L. cv. Xanthi) cells is inhibited by all three solutes. Salt tolerance in Dunaliella parva, which is due to internal formation of glycerol, is accompanied by the adaptation of the activity of the enzyme to elevated glycerol concentrations.     

Ultrastructure and taxonomy of a marine photosynthetic stramenopile Phaeomonas parva gen. et sp. nov. (Pinguiophyceae) with emphasis on the flagellar apparatus architecture

Phaeomonas parva gen. et sp. nov., a marine photosynthetic stramenopile from oceanic water near the Caroline Islands, is described. Cells are naked and spherical to ovoid. The alga is motile with two laterally inserted flagella during the light period, whereas during the dark period, it absorbs the flagella and rounds up. The anterior (immature, No. 2) long flagellum possesses tubular tripartite mastigonemes. The posterior (mature, No. 1) short flagellum is smooth and has autofluorescence at the base. The cupshaped, yellowish‐brown chloroplast occupies the posterior half of the cell, and a pyrenoid occurs in the inner cavity of the cup‐shaped chloroplast. The flagellar apparatus has several unusual features. Two basal plates and a two‐gyred proximal helix in the flagellar transitional region may suggest that P. parva is related to the Pelagophyceae, Dictyochophyceae and Sulcochrysis biplastida, a photosynthetic stramenopile of uncertain taxonomic position. The R3 and R4 roots form a loop that resembles phagotrophic chrysophytes. However, this resemblance is superficial because Phaeomonas is not phagotrophic, its R3 root has a different number of microtubules and its R3 root does not split to form a food‐uptake mouth. Phaeomonas has a ‘bypassing root’, which is found only with the Phaeophyceae, Giraudyopsis stellifera (Chrysomerophyceae), and Ankylochrysis lutea (probably a member of the Pelagophyceae). The taxonomic position of P. parva could not be determined solely from ultrastructural features. However, molecular phylogeny and biochemical analyses (published separately) strongly supported a relationship between P. parva and four other monotypic strameno‐piles, Glossomastix, Pinguiochrysis, Pinguiococcus and Polypodochrysis. Although these algae are morphologically distinct, they have unusually high percentages of polyunsaturated fatty acids, especially eicosapentoic acid. This unusual assemblage of stramenopiles is classified in a new class, the Pinguiophyceae (published separately), and P. parva is its only biflagellate member.     

THE GLYCEROL PERMEABILITY OF THE PLASMALEMMA OF THE HALOTOLERANT GREEN ALGA DUNALIELLA PARVA (VOLVOCALES)1

The glycerol permeability of the plasmalemma of the green alga Dunaliella parva Lerche was investigated by efflux studies with labelled glycerol, by enzymatic determination of glycerol leakage, and the determination of the reflection coefficient from osmotically induced volume changes (zero flow method). All results indicate that the plasmalemma of D. parva does not exhibit a special low permeability towards glycerol as would be expected from a glycerol accumulating alga. Rather, significant amounts of glycerol diffuse continuously into the medium following the glycerol concentration gradient between the cells and the medium. Efflux rates vary between 0.1 and 2 μmoles glycerol·mg^?1 chlorophyll·h^?1 depending on the external NaCl concentration. After one day up to 25% of the total glycerol of the algal suspension was found in the medium. Within 10 days this value can increase to 60%, depending on the growth constant of the culture. The reflection coefficient σ was determined to be 0.87, the permeability coefficient 2800 × 10^?11 m·sec^?1. To maintain a proper endogenous glycerol level corresponding to the external osmotic pressure, glycerol efflux in D. parva has to be balanced by a continuous synthesis of glycerol. D. parva follows the strategy of “glycerol efflux tolerance” instead of “glycerol efflux avoidance”. The alga has to pay the energetic costs of this strategy of tolerance.     

PHYLOGENETIC RELATIONSHIP AMONG VARIOUS STRAINS OF DUNALIELLA (CHLOROPHYCEAE) BASED ON NUCLEAR ITS rDNA SEQUENCES

Phylogenetic analyses of 15 strains representing 8 taxa of Dunaliella (D. salina, D. bardawil, D. pseudosalina, D. tertiolecta, D. parva, D. viridis, D. peircei, and D. lateralis) belonging to both subgenera and all sections of the genus were carried out using the sequences of the nuclear rDNA spacers (internal transcribed spacer [ITS-1 + ITS-2]). The ITS data agreed with the traditional data in that D. lateralis (from subgenus Pascheria) is only distantly related to the seven taxa of the subgenus Dunaliella. The ITS data also supported the monophyly of the subgenus Dunaliella. Within the subgenus Dunaliella, sequence data resolved five phylogenetic groups; some isolates of D. parva and D. salina separated into different clades containing other species. For example, D. parva UTEX 1983 (section Dunaliella) grouped with D. viridis CONC 002 (section Virides); the former has more nucleotides in common with D. viridis (93.2% similarity) than to its conspecifics (85.6% similarity). Likewise, the strains of D. parva CCMP 362 and CCAP 19 / 9 (section Dunaliella), the three strains of D. tertiolecta (section Tertiolectae), and the one strain of D. peircei (section Peirceinae) formed a strong phylogenetic clade (99%–100% support). Dunaliella salina UTEX 200 is more closely related to D. pseudosalina CONC 010 than to its conspecifics (95% similarity), even though the two taxa differ markedly physiologically. The results revealed that D. parva UTEX 1983 has been misidentified and should be renamed as D. viridis. Similarly, the strains of D. parva CCAP 19 / 9 and CCMP 362 and the strain UTEX 2192 of D. peircei should be renamed as D. tertiolecta. More physiological and molecular work needs to be done to elucidate the correct taxonomic position of D. salina UTEX 200 and D. pseudosalina CONC 010. Finally, the high ITS sequence variability found among the various strains of D. salina underlines the importance of further work to elucidate the species status in this complex taxon.