The Calicotyle conundrum: do molecules reveal more than morphology?

Partial large subunit 28S rDNA sequences were obtained for specimens of Calicotyle (Monogenea: Monocotylidae) from eight different host species distributed worldwide to test the validity of some species and to address the question of host-specificity in others. Sequences obtained for Calicotyle specimens identified as C. kroyeri based on morphological methods from the type-host Raja radiata (Rajidae) and an additional host R. clavata, both from the North Sea, were identical. However, `C. kroyeri' from the cloaca of R. naevus from Tunisia, Raja sp. A from Tasmania and R. radula from Tunisia differed from C. kroyeri from R. radiata by five (0.51%), 21 (2.13%) and 39 (3.96%) base pairs, respectively, over 984 sites. Therefore, it is likely that the specimens from Raja sp. A, R. radula and perhaps even from R. naevus are not C. kroyeri. Molecular results determined that the calicotylines from the cloaca of Urolophus cruciatus and U. paucimaculatus (Urolophidae) from southern Tasmania identified previously as C. urolophi are indeed identical. Large subunit 28S rDNA sequences of C. palombi and C. stossichi collected from the cloaca and rectal gland, respectively of Mustelus mustelus (Triakidae) from the coast of Tunisia differ sufficiently for these calicotylines to be considered separate and valid species. Our results indicate that some species of Calicotyle are not strictly host-specific, but that C. kroyeri may not be as widely distributed in rajids as was believed previously. Calicotyle specimens from rajids must be re-examined critically to determine whether there are morphological differences indicative of specific differences that may have been overlooked previously.     

A new species of Calicotyle Diesing, 1850 (Monogenea: Monocotylidae) from the shortspine spurdog Squalus?mitsukurii Jordan & Snyder and the synonymy of Gymnocalicotyle Nybelin, 1941 with this genus

Calicotyle japonica n. sp., collected from the uterus, rectal gland, archinephric duct and cloaca of the shortspine spurdog Squalus mitsukurii Jordan & Snyder (Squaliformes) off the Pacific coast of Japan, is described. The new species can be distinguished from C. inermis Woolcock, 1936 by the shape of the male copulatory organ; in C. japonica this is directed anteriorly, is sharply bent in the middle and then increases in width toward the tip, whereas it is long, coiled and uniform in width throughout its entire length in C. inermis. Furthermore, the intestinal caeca have many irregular diverticula on both sides and the vaginal apertures are at the level of the common genital pore in C. japonica, whereas in C. inermis the intestine is smooth and the vaginae open at the level of the o?type. A phylogeny constructed using LSU rDNA data indicates that the new species is grouped with other Calicotyle species; based on this, Gymnocalicotyle Nybelin, 1941 is synonymised with Calicotyle Diesing, 1850. Calicotyle is divided into two major clades, with the new species being grouped with Calicotyle species infecting sharks and the second clade consisting of Calicotyle spp. infecting rays. This suggests that the loss of the hamuli in C. japonica and C. inermis, both parasites of sharks, is a comparatively recent event in the evolution of the genus.     

Calicotyle hydrolagi n. sp. (Monogenea: Monocotylidae) infecting the deep-sea Eastern Pacific black ghost shark Hydrolagus melanophasma from the Atacama Trench,with comments on host specificity of Calicotyle spp.

We describe Calicotyle hydrolagi n. sp. (Monogenea: Monocotylidae) infecting the cloaca of deep-water Eastern Pacific black ghost sharks, Hydrolagus melanophasma captured as bycatch at a local fishery for Patagonian toothfish Dissostichus eleginoides, (Nototheniidae) in the Atacama Trench using morphological and nucleotide (LSU rRNA and SSU rRNA) data. This new species is differentiated from its congeners by a number of characters, including the absence of a cecal diverticula, the size and shape of the male copulatory organ and the shape of the vagina, as well as by differences in molecular data (SSU rRNA and LSU rRNA). The suitability of some sclerotized structures such as the male copulatory organ (MCO) as a taxonomic character is discussed; specifically, we found that the relationship between MCO and total length exhibit different trends in members of Calicotyle isolated from sharks, skates and chimaeras. Additional efforts to obtain sample of Calicotyle species and further molecular studies based on ribosomal and mitochondrial genes are necessary to clarify the degree of host specificity in this genus. Additionally, this is the first report of a member of Calicotyle to be reported in the Southeastern Pacific Ocean.     

Calicotyle australiensis n. sp. and Calicotyle sp. (Monogenea,Monopisthocotylea) from the rectum and rectal glands,and Rugogaster hydrolagi Schell, 1973 (Trematoda,Aspidogastrea) from the rectal glands of holocephalans off the coast of southeastern Australia

Calicotyle australiensis n. sp. from Chimaera sp., caught off the coast of New South Wales, is described. It differs from the other species of the genus in the combination of the following characteristics: length of the penis-tube, absence of medial diverticula of the caeca and presence of hamuli. Calicotyle sp. from Rhinochimaera pacifica and Rugogaster hydrolagi from Chimaera sp. caught at the same locality are reported and illustrated.

     

Calicotyle australiensis n. sp. and Calicotyle sp. (Monogenea,Monopisthocotylea) from the rectum and rectal glands,and Rugogaster hydrolagi Schell, 1973 (Trematoda,Aspidogastrea) from the rectal glands of holocephalans off the coast of southeastern Australia

Calicotyle australiensis n. sp. from Chimaera sp., caught off the coast of New South Wales, is described. It differs from the other species of the genus in the combination of the following characteristics: length of the penis-tube, absence of medial diverticula of the caeca and presence of hamuli. Calicotyle sp. from Rhinochimaera pacifica and Rugogaster hydrolagi from Chimaera sp. caught at the same locality are reported and illustrated.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores. II. Six New Species from Japanese Shrew Moles (Talpidae)1

ABSTRACT. Thirty-eight of 51 (74.5%) shrew moles collected in Japan were infected with from one to four species of Eimeria and/or Isospora including six of six Dymecodon pilirostris and 32 of 45 (71.1%) Urotrichus talpoides. Four eimerians and two isosporans were identified and all are described as new species. Sporulated oocysts of Eimeria amorphospora n. sp. were subspheroid/ellipsoid, 21.1 × 17.9(18-25 × 16-21) μm. Sporocysts were amorphous, gelatinoid envelopes 20.3 × 7.5 (17–24 × 7–9) μm. Sporozoites were enclosed together within a membrane in each sporocyst. This species was found in 9 of 45 (20%) U. talpoides. Sporulated oocysts of Eimeria gonocilia n. sp. were subspheroid/ellipsoid, 28.8 × 24.4 (25–30 × 21–28) μm; a highly ornate outer oocyst wall gave the appearance of a ciliated ball. Sporocysts ovoid, pointed at both ends, were 17.0 × 9.0 (15–19 × 7–11) μm; this species was found in 4 of 45 (8.9%) U. talpoides. Sporulated oocysts of Eimeria talpoidei n. sp. were asymmetrical ovoid, 20.6 × 13.3 (18–23 × 12–15) μm, with sporocysts lacrimiform, 12.0 × 5.8 (10–14 × 5–7) μm. This species was found in 7 of 45 (15.6%) U. talpoides. Sporulated oocysts of Eimeria honshuensis n. sp. were ellipsoid, 15.5 × 11.4 (13–18 × 10–13) μm, with sporocysts ovoid, 9.1 × 5.2 (8–10 × 4–6) μm. This species was found in 10 of 45 (22.2%) U. talpoides and in 5 of 6 (83.3%) D. pilirostris. Sporulated oocysts of Isospora dymecodi n. sp. were subspheroid/ellipsoid, 15.8 × 12.6 (13–17 × 11–13) μm, with sporocysts ellipsoid, 10.9 × 6.9 (10–13 × 6–8). This species was found in six of six D. pilirostris. Sporulated oocysts of Isospora urotrichi n. sp. were spheroid/subspheroid, 13.4 × 12.4 (11–16 × 9–14) μm, with sporocysts ovoid, 9.2 × 6.3 (8–11 × 5–7) μm. This species was found in 27 of 45 (60%) U. talpoides. Only 14 of 38 (36.8%) infected hosts (one D. pilirostris, 13 U. talpoides) were seen to be naturally infected with only one coccidian species when sampled.     

Ulva (Chlorophyta,Ulvales) Biodiversity in the North Adriatic Sea (Mediterranean,Italy): Cryptic Species and New Introductions

Ulva Linnaeus (Ulvophyceae, Ulvales) is a genus of green algae widespread in different aquatic environments. Members of this genus show a very simple morphology and a certain degree of phenotypic plasticity, heavily influenced by environmental conditions, making difficult the delineation of species by morphological features alone. Most studies dealing with Ulva biodiversity in Mediterranean waters have been based only on morphological characters and a modern taxonomic revision of this genus in the Mediterranean is not available. We report here the results of an investigation on the diversity of Ulva in the North Adriatic Sea based on molecular analyses. Collections from three areas, two of which subject to intense shipping traffic, were examined, as well as historical collections of Ulva stored in the Herbarium Patavinum of the University of Padova, Italy. Molecular analyses based on partial sequences of the rbcL and tufA genes revealed the presence of six different species, often with overlapping morphologies: U. californica Wille, U. flexuosa Wulfen, U. rigida C. Agardh, U. compressa Linnaeus, U. pertusa Kjellman, and one probable new taxon. U. californica is a new record for the Mediterranean and U. pertusa is a new record for the Adriatic. Partial sequences obtained from historical collections show that most of the old specimens are referable to U. rigida. No specimens referable to the two alien species were found among the old herbarium specimens. The results indicate that the number of introduced seaweed species and their impact on Mediterranean communities have been underestimated, due to the difficulties in species identification of morphologically simple taxa as Ulva.     

Temperature requirements for the growth and maturation of the gametophytes of Undaria pinnatifida and U. undarioides (Laminariales, Phaeophyceae)

Gametophytes of two Undaria species, U. pinnatifida and U. undarioides (Laminariales, Phaeophyceae), were studied to determine their water temperature requirements in order to understand their different distributions in Mie Prefecture, Japan. The optimal temperature for growth was 20°C for gametophytes of both species, and the upper critical temperature for growth was also the same for both species at 28°C. Therefore, the optimal and critical temperatures for growth of the gametophytes are not the main factors determining distribution. The optimal temperature for maturation of U. pinnatifida was approximately 10–15°C, whereas it was closer to 20–21°C for U. undarioides, a difference between these species of at least 5°C. In autumn and early winter, the seawater temperature at the mouth of Ise Bay, where U. pinnatifida is distributed, ranges from 21.6°C (October) to 12.7°C (December), and off Hamajima, where U. undarioides is found, the range is from 22.7°C (October) to 19.1°C (December). The seawater temperatures from October to December, which is the maturation season for the gametophytes, agreed well with the optimal temperature requirements for maturation of the gametophytes of both species. Thus the difference in the maturation temperature range of the gametophytes is a major factor determining distribution of these Undaria species along the Japanese coast.     

Chrysosphaerella (Pseudochrysosphaerella)solitaria,spec. nova (Chrysophyceae)

A new unicellular species of the genusChrysosphaerella (Chrysophyceae) was found in fresh-water ponds in Switzerland, Japan, and the U.S.A. It is described asC. solitaria. The genus is divided into two subgenera:Chrysosphaerella, comprising the colonial species, andPseudochrysosphaerella, the unicellular ones.     

MOLECULAR ASSESSMENT OF ULVA SPP. (ULVOPHYCEAE,CHLOROPHYTA) IN THE HAWAIIAN ISLANDS1

Sequences of the nuclear internal transcribed spacer 1 (ITS1) region and the chloroplast rbcL gene were obtained from 86 specimens of Ulva (including “Enteromorpha”) from five of the main Hawaiian Islands. These 86 specimens were divided into 11 operational taxonomic units (OTUs) based on analyses of primary sequence data and comparisons of ITS1 secondary structure. Of the 11 OTUs, six have not previously been reported from anywhere in the world. Only three represented exact sequence matches to named species (Ulva lactuca L., syn. U. fasciata Delile; U. ohnoi Hiraoka et Shimada); two others represented exact sequence matches to unnamed species from Japan and New Zealand. Of the 12 species names currently in use for Hawaiian Ulva, only one, U. lactuca (as U. fasciata), was substantiated. General morphology of the specimens did not always correspond with molecular OTUs; for example, reticulate thallus morphology, previously considered diagnostic for the species U. reticulata Forssk., was expressed in thalli assigned to U. ohnoi and to one of the novel OTUs. This finding confirms a number of recent studies and provides further support for a molecular species concept for Ulva. These results suggest that Ulva populations in tropical and subtropical regions consist of species that are largely unique to these areas, for which the application of names based on types from temperate and boreal European and North American waters is inappropriate. Ulva ohnoi, a “green tide” species, is reported from Hawaii for the first time.     

ACACIA CONSTRICTA (FABACEAE: MIMOSOIDEAE) AND RELATED SPECIES FROM THE SOUTHWESTERN U.S. AND MEXICO

Vegetative, fruit, and floral characters of more than 200 herbarium specimens of Acacia constricta and closely related species from the Southwestern U.S. and Mexico were scored and analyzed by principal components analysis (PCA) and stepwise discriminant analysis (SDA). The results show that all of the 5 species of this group are distinct and form nonoverlapping groups in plots of the first three principal components. All taxa were found to be quite homogeneous as indicated by the tight clusters formed in the PCA plots. Acacia pacensis, whose relationship to the species of this group has been questioned, is confirmed to be more closely related to A. farnesiana and related species. Some specimens with a combination of characters from A. constricta, A. neovernicosa, and A. schottii were positioned intermediate to the putative parental species groups in the PCA and SDA plots, suggesting hybridization among these taxa.     

Molecular assessment of Ulva (Ulvales,Chlorophyta) diversity in Vietnam including the new species U. vietnamensis

Species diversity of Ulva in Vietnam was investigated using three commonly used genetic markers, the nuclear encoded rDNA ITS region and the plastid encoded rbcL and tufA genes. Single locus species delimitation methods, complemented with morphological and ecological information resulted in the delimitation of 19 species. This diversity is largely incongruent with the traditional understanding of Ulva diversity in Vietnam. Only four species identified in this study, U. lactuca, U. reticulata, U. spinulosa, and U. flexuosa, have been previously reported, and seven species, U. ohnoi, U. tepida, U. chaugulii, U. kraftiorum, U. meridionalis, U. limnetica, and U. aragoënsis, are recorded for the first time from Vietnam. Seven genetic clusters could not be associated with species names with certainty. A new species, U. vietnamensis, is described from marine to brackish coastal areas from southern Vietnam based on its morphological and molecular distinctiveness from the currently known Ulva species. A comparison with recent molecular-based studies of Ulva diversity showed that species composition in Vietnam is similar to that of adjacent countries, including Japan, China, as well as Australia. Our study emphasizes the importance of molecular data in the assessment of Ulva diversity, and indicates that a lot of diversity may still remain to be discovered, especially in tropical regions.     

Behavioural adaptations of the fiddler crabs Uca vocans borealis (crane) and Uca lactea lactea (De Haan) for coexistence on an intertidal shore

The distribution and coexistence of the fiddler crabs U. vocans borealis and U. lactea lactea was investigated in the upper shore of a sandy beach with respect to particle size, water content and total organic carbon content of the sediments. A clear segregation of habitats between the two species was apparent. U. lactea lactea were only found in sediments with significantly lower total organic carbon content. U. lactea lactea had a higher ratio for the size of third maxilliped to body size than U. vocans borealis. There was no statistical difference in the median sediment particle size of the habitats where both species were found on the study shore. U. lactea lactea was larger on shores where U. vocans borealis were absent. Smaller U. vocans borealis individuals occupied sediments with higher water content and finer particles. Smaller individuals of U. vocans borealis also possessed fewer spoon-tipped setae on second maxilliped than their larger conspecifics, which were found mostly on coarser sediments. U. lactea lactea spent significantly more time on the surface than in the burrow during low tide when compared with U. vocans borealis. Both Uca species exhibited similar desiccation resistance. U. lactea lactea tends to keep its frontal region in close contact with sediments to maintain moisture when being exposed to air.     

Cryptic diversity of Ulva (Ulvales,Chlorophyta) in the Great Bay Estuarine System (Atlantic USA): introduced and indigenous distromatic species

Distromatic foliose blades of the algal genus Ulva are notoriously difficult to identify due to their simple morphologies and few diagnostic characteristics that often exhibit intraspecific variation and interspecific overlap. Hence, species differentiation is difficult and diversity estimates are often inaccurate. Two major goals of this study were to assess the diversity of distromatic Ulva spp. in the Great Bay Estuarine System (GBES) of New Hampshire and Maine, USA, and to compare historical and present day records of these species. Molecular analysis (using ITS sequences) of field-collected specimens revealed four distinct taxa: Ulva lactuca, U. rigida, U. compressa, and U. pertusa. Prior to molecular screening, Ulva lactuca was the only distromatic Ulva species reported for the GBES. Ulva pertusa and the foliose form of U. compressa are newly recorded for the Northwest Atlantic, and the range of U. rigida has been extended. Molecular analysis of historical herbarium voucher specimens indicates that U. rigida, U. pertusa, and the foliose form of U. compressa have been present in the GBES since at least 1966, 1967, and 1972, respectively. The distromatic morphotype of U. compressa is found only in low salinity areas, which suggests that salinity may influence its morphological development. Molecular and morphological evaluations are critical if we are to distinguish between cryptic taxa, accurately assess biodiversity, and effectively monitor the spread of non-indigenous macroalgae.     

Fiddler crab (Crustacea: Ocypodidae) distribution and the relationship between habitat occupancy and mouth appendages

Sediment characteristics, especially grain size, are usually considered the most important variables affecting Uca distribution, mainly due to its close relationship with mouth appendage morphology. The aim of this study was to verify, from an assemblage perspective, if sediment is the most important variable affecting Uca species distribution, and if mouth appendage morphology (setae type and curvature) would be related to habitat occupancy. Niche metrics and null model approaches were used to assess and test the hypothesis. The relevance of spoon-tipped setae curvature to Uca distribution was verified for the first time. A fragmented mangrove area was divided into seven subareas, and sampling of crabs and environmental variables took place in June and November 2010. Of 10 species recorded for Brazil, seven were found in the study area: U. burgersi, U. cumulanta, U. leptodactyla, U. maracoani, U. rapax, U. thayeri and U. uruguayensis. Multivariate analysis showed that sediment grain size and the presence of vegetation were the most important variables explaining distribution, reinforcing results commonly obtained by univariate approaches. The overlap of habitat occupancy was generally low and no relationship between mouth appendages was found with breadth and overlap measures. Contrary to predictions, most non-random overlap values were lower than expected by chance, suggesting that interspecific competition might influence species distribution. Also, variables such as the presence of vegetation are important and influence crab distribution, limiting the potential distribution that would be predicted by mouth adaptations alone. Thus, the use of these adaptations as surrogates of fiddler crab distribution is not recommended.     

Karyotype relationships among four South American species of Urvillea (Sapindaceae: Paullinieae)

A cytogenetic study was conducted on four species of the genus Urvillea (Sapindaceae, Paullinieae): U. chacoensis Hunz., U. filipes Radlk. and U. ulmacea Kunth of the Urvillea section and U. laevis Radlk. of the Stenelytron section. The chromosome numbers in U. chacoensis (2n = 22) and U. laevis (2n = 24) were confirmed, and new chromosome numbers are reported for U. filipes with 2n = 22 and U. ulmacea with 2n = 88. Additionally, data on interphase nuclear structure, chromosome banding patterns (C-Giemsa and C-CMA₃/DAPI) and FISH with rDNA probes are also presented. The distribution of AT- and GC-rich regions and the physical mapping of ribosomal genes (45S and 5S rDNA sites) were established for the first time in these Urvillea species. Sections of Urvillea are cytogenetically differentiated according to basic chromosome number, where x = 11 in the section Urvillea and x = 12 in the section Stenelytron. This first section displayed an important karyotypic feature, the occurrence of large AT- and GC-rich bands at terminal chromosomal regions. The Urvillea section showed polyploidy and its species were differentiated by their banding patterns. Urvillea chacoensis showed several terminal AT-rich bands, while terminal AT- and GC-rich bands were both found in U. ulmacea. However, the section Stenelytron did not exhibit this banding pattern. The 45S rDNA sites appeared always associated with GC-rich regions and they were numerically variable among species, being located or not the same chromosome 5S rDNA sites. Variation in the repetitive DNA distribution and their role in karyotype differentiation among these Urvillea species are discussed.     

Molecular identification and distribution of leatherjackets (Diptera: Tipulidae) in U.K. agricultural grassland

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REAPPRAISAL OF UTRICULARIA INFLATA AND U. RADIATA (LENTIBULARIACEAE)

Reinert , Grady , W., and R. K. Godfrey . (Florida State U., Tallahassee.) Reappraisal of Utricularia inflata and U. radiata (Lentibulariaceae). Amer. Jour. Bot. 49(3): 213–220. Illus. 1962.—Utricularia inflata and U. radiata, interpreted either as separate species or 2 varieties of a single species by previous investigators, are shown to be specifically distinct on the basis of 5 hitherto-neglected, morphological characters, 4 of which are vegetative in nature, and on the basis of 3 reinvestigated quantitative criteria. The particular attention devoted to the vegetative bodies of the 2 plants led to the discovery, in U. inflata, of tuber formation, a phenomenon known to occur in Utricularias in general but unreported for North American species. Support for interpreting U. inflata and U. radiata as species resides in the distribution patterns. Although both occur on the outer coastal plain from southern Alabama to southern Delaware, the 2 taxa are easily distinguishable and exhibit no intergradation. The range of U. radiata is much broader; it extends westward to eastern Texas northeastward to Nova Scotia, and is disjunct inland in western Tennessee, western Virginia and in northwestern Indiana.     

Heterotrissocladius boltoni sp.n., a new orthoclad from vernal pools and streams in Ohio,U.S.A. (Diptera: Chironomidae)

Heterotrissocladius boltoni sp.n. from Ohio, U.S.A., is described in both sexes and all stages. The species belongs in themarcidus group and is found in temporary streams and pools