THE BIOMPHALARIA HAVANENSIS COMPLEX (GASTROPODA: PLANORBIDAE) IN CUBA: A MORPHOLOGICAL AND GENETIC STUDY

Conchological, anatomical and genetic characteristics werecompared among several populations of the Biomphalaria havanensis complexfrom Cuba and other localities in the Caribbean region. Two morphometricmeasures (height and diameter) distinguished two closely similarmorphs of the shell. The reproductive system also separated thesemorphs, particularly upon comparison of sizes of the penis sheathand the preputium. The two morphological groups differed in9 to 12 fixed alleles according to population. Further allozymicdata analyzed in the present work confirmed this difference.The results strongly support the assumption that the B. havanensis complexincludes two distinct species: B. havanensis, for which thetype locality is the swampy area near the vicinity of Havana,Cuba and Biomphalaria sp. a species which commonly occurs inCuba and in Dominican Republic. The taxonomic identity of Biomphalariasp. is discussed. (Received 28 January 2000; accepted 15 August 2000)     

Experimental deep-sea deployments reveal diverse Northeast Pacific wood-boring bivalves of Xylophagainae (Myoida: Pholadidae)

Only a single species of deep-sea wood boring bivalve has beenknown off the forested northwest coast of North America, althoughthe Xylophagainae are so diverse in other areas that up to fivespecies occur in a given length of wood. To determine whetheradditional species were present in the Northeast Pacific andif so, how they coexist, lengths of wood were experimentallydeployed on heavily sedimented sites on the Cascadia Basin andEscanaba Trough, non-hydrothermally active basalt on Gorda Ridgeand Axial Volcano, and an isolated sediment pond within theaxis of Juan de Fuca Ridge, at depths of between 1,530 and 3,232 m.All locations were between 41°and 48°N, at least 240 kmoff the North American coast. Six previously unknown species,Xylophaga corona n. sp., X. zierenbergi n. sp., X. heterosiphonn. sp., X. oregona n. sp., X. microchira n. sp. and Xylopholascrooki n. sp., were collected and are described here. Dominantspecies differed in recoveries made after 10 and 24 months atJuan de Fuca Ridge and Cascadia Basin localities. Xylophagamicrochira n. sp. dominated four of five deployments recoveredwithin 12 months, but its abundance declined by 24 months, beinglargely replaced on Juan de Fuca Ridge by X. oregona n. sp.,a species known only from this Ridge and from an incidentalcollection on the Nootka Fracture Zone. At Cascadia Basin sites,the abundances of X. heterosiphon n. sp. and of what are likelyto be predatory polyclad flatworms increased as that of X. microchiran. sp. declined. Xylophaga microchira n. sp. is hypothesizedto be specialized for early colonization, but to be competitivelyinferior to X. oregona n. sp., and more vulnerable to predatorsthan X. heterosiphon n. sp. The dominance of competitively superiorand more predator-resistant species in older deployments arguesthat biotic interactions contribute to coexistence of multiplespecies of wood-borers despite direct competition among them. (Received 2 February 2007; accepted 16 August 2007)     

Paradiscogaster flindersi and P. oxleyi n. sp. (Digenea: Faustulidae): overlapping host and geographical distributions in corallivore chaetodontid fishes in the tropical Indo-west Pacific

A total of 2,868 individuals of 47 species of chaetodontids were examined for faustulids at seven major localities in the Tropical Indo-West Pacific (TIWP). Combined morphological and molecular analyses allowed us to describe Paradiscogaster oxleyi n. sp. from three localities in the TIWP and in three host species, Chaetodon lunulatus Quoy & Gaimard (type-host), C. ornatissimus Cuvier and C. meyeri Bloch & Schneider. Molecular analysis of the ITS2 region of rDNA from two host species and three localities supports the morphology-based conclusion that P. oxleyi n. sp. is the same species at the three localities. Paradiscogaster flindersi Bray, Cribb & Barker, 1994 is reported from three new localities in the TIWP and is now known from 13 chaetodontid species. Sequences from samples consistent with P. flindersi differed from those from P. oxleyi n. sp. in 11–12 base pairs. The host ranges of the two species overlap broadly. Neither species was found in French Polynesia but both were found at Swain Reefs on the Great Barrier Reef. Only one of the two species was found at each of the five other sites. Both species occur almost exclusively in specialist corallivores allowing the inference that the metacercariae occur in corals. Finally, a key to the species of Paradiscogaster is provided.     

Morphological and molecular data reveal a new species of Neoechinorhynchus (Acanthocephala: Neoechinorhynchidae) from Dormitator maculatus in the Gulf of Mexico

Neoechinorhynchus (Neoechinorhynchus) mexicoensis sp. n. is described from the intestine of Dormitator maculatus (Bloch 1792) collected in 5 coastal localities from the Gulf of Mexico. The new species is mainly distinguished from the other 33 described species of Neoechinorhynchus from the Americas associated with freshwater, marine and brackish fishes by having smaller middle and posterior hooks and possessing a small proboscis with three rows of six hooks each, apical hooks longer than other hooks and extending to the same level as the posterior hooks, 1 giant nucleus in the ventral body wall and females with eggs longer than other congeneric species. Sequences of the internal transcribed spacer (ITS) and the large subunit (LSU) of ribosomal DNA including the domain D2 + D3 were used independently to corroborate the morphological distinction among the new species and other congeneric species associated with freshwater and brackish water fish from Mexico. The genetic divergence estimated among congeneric species ranged from 7.34 to 44% for ITS and from 1.65 to 32.9% for LSU. Maximum likelihood and Bayesian inference analyses with each dataset showed that the 25 specimens analyzed from 5 localities of the coast of the Gulf of Mexico parasitizing D. maculatus represent an independent clade with strong bootstrap support and posterior probabilities. The morphological evidence, plus the monophyly in the phylogenetic analyses, indicates that the acanthocephalans collected from intestine of D. maculatus from the Gulf of Mexico represent a new species, herein named N. (N.) mexicoensis sp. n.     

Morphological and molecular differentiation of two new species of Pseudoacanthocephalus Petrochenko, 1958 (Acanthocephala: Echinorhynchidae) from amphibians and reptiles in the Philippines, with identification key for the genus

The genus Pseudoacanthocephalus Petrochenko, 1958 currently includes 14 species of acanthocephalans parasitic in amphibians and reptiles worldwide. This work describes two new species of Pseudoacanthocephalus from amphibians and reptiles collected in several localities on Luzon Island, Philippines. Pseudoacanthocephalus nickoli n. sp. was found in two species of frogs, Rana luzonensis Boulenger and Rana similis (Günther), and Pseudoacanthocephalus smalesi n. sp. was found in a scincid lizard, Sphenomorphus abdictus Brown & Alcala. Differential diagnoses of the two new species of Pseudoacanthocephalus from their congeners are provided. Comparative analysis of nuclear ribosomal rRNA sequences encompassing the 3′ end of 18S nuclear rDNA gene, internal transcribed spacer region (ITS1+5.8S+ITS2), and 5′ end of the 28S gene strongly corroborated the morphological evidence and demonstrated significant differences between the two new species as well as between these species and closely related species from continental China and Vietnam. No intraspecific sequence variability was detected among different individuals representing each of the examined species. This is the first report of Pseudoacanthocephalus in the Philippines. A key to known species of Pseudoacanthocephalus is provided.     

FOUR NEW SPECIES OF TAMBJA BURN, 1962 (NUDIBRANCHIA: POLYCERIDAE) FROM THE INDO-PACIFIC

The genus Tambja previously included 24 described species. Fournew species, T. tentaculata n. sp., T. gabrielae n. sp., T.zulu n. sp. and Tambja victoriae n. sp., from the Indo-Pacificare described. Tambja tentaculata n. sp., from Guam, is theonly known species in the genus with well developed, dorsolaterallygrooved, oral tentacles. Its inner lateral teeth have a bifidinner cusp with two long, sharp denticles. The oral tentaclesof T. tentaculata are more typical of Roboastra species, whilethe shape of the inner lateral teeth is more typical of Tambja.Nevertheless, the arrangement of the two cusps of the innerlateral teeth and the presence of a rachidian tooth withoutdenticles and with a central notch at the anterior edge, typicalof the species of the genus Tambja, suggest the placement withinthis genus. Tambja gabrielae n. sp., from Indonesia and PapuaNew Guinea, has dark green to dark brown ground colour withbright yellow patches scattered on the body. Tambja zulu n.sp. from Durban, South Africa, is characterized by a black groundcolour with slender yellow longitudinal lines. Tambja victoriaen. sp. is a new species from Papua New Guinea and Australiathat has frequently been misidentified as Roboastra arika, characterizedby its blue body colour and yellow lines. The four species aredistinguishable based on differences in body coloration, ofcharacters of the radula and of the reproductive system. Anoverview on distinguishing features of all known Indo-PacificTambja species is presented. (Received 21 June 2004; accepted 20 January 2005)     

Description of two new species of Glypthelmins Stafford, 1905 (Digenea: Macroderoididae) in Rana spp. from Mexico, based on morphology and mtDNA and rDNA sequences

Glypthelmins Stafford, 1905 includes 29 putative species commonly found in the intestine and liver of anurans from all over the world but mainly in the Americas. Partial sequences of the cytochrome c oxidase subunit 1 ( cox 1), ribosomal internal transcribed spacer region 2 (ITS2) and the large subunit 28S rDNA gene were obtained and analysed using pairwise distance matrices and parsimony methods in order to characterise the interrelationships between 14 isolates of four nominal species of Glypthelmins recognised on morphological grounds. The highest intra-specific sequence divergence occurred in the cox 1 (18.53%) sequence, followed by that of the ITS2 (5.44%) and 28S (4.63%). Genetic variability was detected between the three isolates originally identified as G. facioi Brenes et al., 1959 from two localities in Mexico and one locality in Costa Rica. Sequence divergence exhibited among these isolates ranged from 10.70 to 11.22%, from 0.48 to 0.97% and from 1.33 to 1.88% for cox 1, ITS2 and 28S, respectively. Phylogenetic analysis combining all three data-sets generated a single most parsimonious tree. The three isolates of G. facioi form a clade, with an isolate collected from frogs in Veracruz State as the sister group to an isolate from Tabasco State + G. facioi from Costa Rica. The information derived from pairwise distance of independent data-sets plus the phylogenetic information indicate that each of the two isolates from Mexico, identified a priori as G. facioi, represent separate species. A re-examination of specimens was carried out and a re-evaluation made of the morphological characters to find reliable differences that had been overlooked. As a consequence, G. brownorumae n. sp. from Tabasco and G. tuxtlasensis n. sp. from Veracruz are described based on molecular and morphological differences.     

Evidence for extensive cryptic speciation in trematodes of butterflyfishes (Chaetodontidae) of the tropical Indo-West Pacific

Molecular data from the cytochrome c oxidase subunit I (cox1) mitochondrial DNA gene and the second internal transcribed spacer (ITS2) nuclear rDNA region were used to test the current morphologically-based taxonomic hypothesis regarding species of Monorchiidae (Hurleytrematoides) from chaetodontid and tetraodontid fishes from six sites in the tropical Indo-West Pacific (TIWP): Heron and Lizard Islands off the Great Barrier Reef (GBR, Australia), Moorea (French Polynesia), New Caledonia, Ningaloo Reef (Australia) and Palau. The 16 morphospecies analysed differed from each other by a minimum of 55 bp (9.1%) over the mitochondrial cox1 and 8 bp (1.6%) over the ITS2 DNA regions. For two species, Hurleytrematoides loi and Hurleytrematoides sasali, specimens from the same host species in sympatry differed at levels comparable to those between pairs of distinct morphospecies for both cox1 and ITS2 sequences. We take this as evidence of the presence of combinations of cryptic species; however, we do not propose new species for these taxa because we lack identified morphological voucher specimens. For seven species, Hurleytrematoides coronatum, Hurleytrematoides deblocki, Hurleytrematoides faliexae, H. loi, Hurleytrematoides morandi, H. sasali and Hurleytrematoides sp. A, samples from some combinations of localities had base pair differences that were equal to or greater than differences between some pairs of distinct morphospecies for one or both cox1 and ITS2 sequences. For three species, H. coronatum, H. loi and H. morandi, one haplotype differed from every other haplotype by more than the morphospecies benchmark. In these cases morphological specimens could not be distinguished by morphology. These data suggest extensive cryptic richness in this genus. For the present we refrain from dividing any of the morphospecies. This is because there is a continuum of levels of intra- and interspecific genetic variation in this system, so that distinguishing the two would be largely arbitrary.     

Morphological and genetic evidence of contemporary intersectional hybridisation in Mediterranean Helichrysum (Asteraceae,Gnaphalieae)

Hybridisation is considered an important evolutionary phenomenon in Gnaphalieae, but contemporary hybridisation has been little explored within the tribe. Here, hybridisation between Helichrysum orientale and Helichrysum stoechas is studied at two different localities in the islands of Crete and Rhodes (Greece). Using three different types of molecular data (AFLP, nrDNA ITS sequences and cpDNA ndhF sequences) and morphological data, the aim is to provide simultaneous and direct comparisons between molecular and morphological variation among the parental species and the studied hybrid populations. AFLP profiles, ITS sequences and morphological data support the existence of hybrids at the two localities studied, shown as morphological and genetic intermediates between the parental species. Chloroplast DNA sequences show that both parental species can act either as pollen donor or as maternal parent. Fertility of hybrids is demonstrated by the viability of seeds produced by hybrids from both localities, and the detection of a backcross specimen to H. orientale. Although there is general congruence of morphological and molecular data, the analysis of morphology and ITS sequences can fail to detect backcross hybrids.     

THE DISTRIBUTION OF SOME ANTARCTIC NUDIBRANCHS (OPISTHOBRANCHIA)

The circumpolar distribution of the four revised species ofdoridacean nudibranchs: Aegires albus, Bathydoris hodgsoni,B. clavigera and Austrodoris kerguelenensis, is demonstrated,taking into consideration the known data from the literatureand new localities from the Atlantic sector of the Antarctic.A. albus and B. hodgsoni are restricted to the High AntarcticZone. A. kerguelenensis is widely distributed in the High Antarcticand Subantarctic Zone, whereas localities of B. clavigera arerestricted to the High Antarctic Zone and the Atlantic sectorof the Subantarctic Zone. (Received 13 November 1986;     

Phylogeny, evolution and biogeography of the Quadrifoliovariinae Yamaguti, 1965 (Digenea: Lecithasteridae)

The Quadrifoliovariinae is revised and three new species of Quadrifoliovarium Yamaguit, 1965 from acanthurid fishes of the genus Naso from waters of the Indo-Pacific are described: Q, maceria n. sp. from N.␣tonganus, N. annulatus, N. fageni and N. brevirostris; Q. simplex n. sp. from N. tonganus and N.␣annulatus; and Q. quattuordecim n. sp. from N. tonganus. Amendments are made to the characterisation of the Quadrifoliovariinae, Quadrifoliovarium, Bilacinia Manter, 1969 and Unilacinia Manter, 1969 in light of observations on type and new material. A molecular phylogeny based on ITS2 and 28S regions of the ribosomal DNA is proposed. The phylogeny suggests that U. asymmetrica is the most basal taxon and Q.␣simplex n. sp. and Q. quattuordecim n. sp. the most derived. Evolution of morphological traits within the Quadrifoliovariinae are discussed in light of the molecular phylogeny. Molecular sequences of the ITS2 rDNA were identical between specimens of Q. pritchardae collected off Exmouth (Indian Ocean), Heron Island and Lizard Island (Western Pacific) and Moorea (far Eastern Indo-Pacific), indicating a broad Indo-Pacific distribution. All members of the subfamily are recorded only from the acanthurid genus Naso, with the exception of B. lobatum (Yamaguti, 1970), which has been recorded from a pomacanthid. The restricted host range of the group is discussed in the light of the phylogeny of the host genus Naso.     

基于rDNA ITS1和ITS2序列的褐飞虱、白背飞虱和灰飞虱的分子鉴定

本研究测定了褐飞虱Nilaparvata lugens、白背飞虱Sogatella furcifera和灰飞虱Laodelphax striatellus的rDNA ITS1和ITS2的序列, 以探讨这3种稻飞虱的分子鉴定方法。3种飞虱的ITS1和ITS2侧翼区（18S, 5.8S和28S）序列相对稳定, 但ITS1和ITS2序列在3种飞虱中变异较大。 ITS1在所分析的438个位点中可变位点达294个, ITS2在分析的403个位点中可变位点为177个。根据3种飞虱rDNA的ITS1和ITS2序列设计了特异性引物, 应用特异性引物对样品进行了PCR扩增, 分析发现3种飞虱ITS1区的特异性引物扩增效果不理想, 而ITS2区的特异性引物可以稳定地扩增出明显的目的DNA条带. 因此, 采用ITS2区的特异性引物可以对3种飞虱进行快速的分子鉴定。     

FRESHWATER SNAILS OF THE BULINUS TRUNCATUS/TROPICUS COMPLEX IN KENYA: TETRAPLOID SPECIES

Studies in recent years on chromosome number in freshwater snailsbelonging to the Bulinus truncatus/tropicus complex demonstratedthe occurrence in Kenya of one diploid species, B. tropicus,and 2 tetraploid species, B. truncatus and B. permembranaceus.Further observations are now described on the morphologies ofthe tetraploid species, and their distributions. Observations were made on B. truncatus from 26 localities (11newly reported) and B. permembranaceus from 26 localities (7newly reported), in respect of chromosome number, egg proteins,enzymes (5 systems), shell (10 variables measured), male genitalsystem (normal or aphallic) and radula (1st lateral tooth, sizeand mesocone shape). B. permembranaceus differs most clearly from B. truncatus inegg proteins, enzyme-loci and in lacking aphallic individuals.Mesocone shape is generally less angular in B. permembranaceus.The shell of B. permembranaceus grows larger, has the columellamore commonly concave and its spire is proportionally higherand more acute. To distinguish these tetraploid species fromthe partly sympatric B. tropicus, observations on chromosomenumber and biochemical characters are necessary. The tetraploids are allopatric, B. truncatus being found rarelyas high as 1900 m altitude, whereas B. permembranaceus occupiesthe altitude range 1940–2760 m. B. truncatus may be expandingits distribution in Kenya in man-made waterbodies, while B.permembranaceus possibly is restricted by adaptation to coolconditions and by interaction with B. tropicus. Differencesbetween B. truncatus and B. permembranaceus, both morphologicaland biochemical, indicate their origins lie in independent episodesof tetraploidy. ^*Member of External Scientific staff, Medical Research Council (Received 31 October 1988; accepted 6 December 1988)     

Calanoid copepods feeding on algae and filamentous cyanobacteria: rates of ingestion, defaecation and effects on trichome length

The relative abilities of freshwater calanoid copepods to grazealgae and filamentous cyanobacteria were examined in three speciesof Boeckella (B.triarticulata, B.hamata, B.dilatata) fed Cydotella,Cryptomonas and two species of Nostoc and Anabaena. Althoughrates of ingestion were highest on Cryptomonas, all the Boeckellaspecies consumed cyanobacteria, some species at similar ratesto Cyclolella. Faecal production on the foods was highest onCydotella and, in B.triarticulata, higher on A.flos-aquae andNostoc sp. 2 than on Cryptomonas. At 1 mg l^–1 of thesecyanobacteria, female B.triarticulata voided 70% by volume ofthe material they ingested as compact faecal pellets. By theirgrazing they also decreased dramatically the trichome lengthsof A.flos-aquae, A.oscillarioides and Nostoc sp. 2, therebypossibly altering the potential susceptibility of these filamentouscyanobacteria to grazing by other zooplankton.     

Characterization and physical mapping of ribosomal RNA gene families in Plantago

• Background and Aims The organization of rRNA genes incultivated Plantago ovata Forsk. and several of its wild allieswas analysed to gain insight into the phylogenetic relationshipsof these species in the genus which includes some 200 species. • Methods Specific primers were designed to amplify theinternal transcribed spacer (ITS1 and ITS2) regions from sevenPlantago species and the resulting fragments were cloned andsequenced. Similarly, using specific primers, the 5S rRNA genesfrom these species were amplified and subsequently cloned. Fluorescencein-situ hybridization (FISH) was used for physical mapping of5S and 45S ribosomal RNA genes. • Results The ITS1 region is 19–29 bp longer thanthe ITS2 in different Plantago species. The 5S rRNA gene-repeatingunit varies in length from 289 to 581 bp. Coding regions arehighly conserved across species, but the non-transcribed spacers(NTS) do not match any database sequences. The clone from thecultivated species P. ovata was used for physical mapping ofthese genes by FISH. Four species have one FISH site while threehave two FISH sites. In P. lanceolata and P. rhodosperma, the5S and 45S (18S-5·8S-25S) sites are coupled. • Conclusions Characterization of 5S and 45S rRNA geneshas indicated a possible origin of P. ovata, the only cultivatedspecies of the genus and also the only species with x = 4, froma species belonging to subgenus Psyllium. Based on the studiesreported here, P. ovata is closest to P. arenaria, althoughon the basis of other data the two species have been placedin different subgenera. FISH mapping can be used as an efficienttool to help determine phylogenetic relationships in the genusPlantago and show the interrelationship between P. lanceolataand P. lagopus.     

Morphological and phylogenetic analyses of <Emphasis Type="Italic">Uromyces appendiculatus</Emphasis> and <Emphasis Type="Italic">U. vignae</Emphasis> on legumes in Japan

Uromyces appendiculatus, inclusive of three varieties, is distinguished from U. vignae primarily by the position of urediniospore germ pores and putative host specificity. However, opinions concerning these morphological and physiological features as taxonomic characters have varied greatly, and distinction of these species has often been confused. To clarify the taxonomy of these two species, morphological features of urediniospores and teliospores of 225 rust fungus specimens on species of Phaseolus, Vigna, Apios, Lablab, and Dunbaria were examined by light microscopy and scanning electron microscopy. Forty-five specimens were subjected to molecular phylogenetic analyses. As a result, the position of germ pores in urediniospores and the teliospore-wall thickness were considered as good characters to separate three morphological groups. In molecular analyses, the specimens fell into two and three clades based on the nucleotide sequence at D1/D2 domain of LSU rDNA and ITS regions, respectively. One of the D1/D2 clades corresponded to one morphological group whereas another D1/D2 clade included two other morphological groups. In contrast, each of the three ITS clades corresponded to a separate morphological group. Neither morphological groups nor molecular clades were host limited. It is suggested that the three morphological groups that corresponded to three distinct ITS clades constitute distinct species.Contribution no. 186 from the Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

Cardicola aurata sp. n. (Digenea: Sanguinicolidae) from Mediterranean Sparus aurata L. (Teleostei: Sparidae) and its unexpected phylogenetic relationship with Paradeontacylix McIntosh, 1934

A new sanguinicolid trematode, Cardicola aurata sp. n., is described from gilthead seabream Sparus aurata L., from off the Spanish Mediterranean coast. The morphology of C. aurata sp. n. generally agrees with the diagnosis of the genus, however, in contrast to all other reported Cardicola spp. the male pore is located sub-medially at the posterior end of the body instead of sinistrally before the posterior end of the body. Based on a comparison of the morphology as well as partial 28S and ITS2 rDNA sequence data from the present species with that from closely related species, it was decided to emend the diagnosis of Cardicola rather than create a new genus, as the aberrant position of the male pore is likely to be an autapomorphy. The phylogenetic analyses revealed a close relationship between Cardicola and Paradeontacylix, two genera with considerable morphological differences; C. aurata sp. n. occupies a position intermediate to these genera. Thus, a morphological comparison of Cardicola, Paradeontacylix and Braya, a genus which is morphologically similar to Cardicola but clusters basal to the Cardicola/Paradeontacylix clade, was conducted. The results of this comparison showed that despite large differences with regard to body shape, the organisation of the internal organs is very similar in species of Cardicola and Paradeontacylix. The synopsis of morphological data and molecular phylogeny allows for interpretations regarding the importance of different morphological features for the phylogenetic inference of the Sanguinicolidae.