On the crustacean hosts of larval acanthocephalan parasites of fishes from a coastal lagoon of River Po Delta

A total of 4664 crustaceans of 2 species of gammarids (Gammarus insensibilis Stock, G. aequicauda Martynon) were collected and examined in search of larval helminth parasites. Larvae of Telosentis exiguus Von Linstow 1901 (Acanthocephala) were found in 6 out of 2806 G. insensibilis so far examined. This is the first record of an intermediate host for T. exiguus, of which a definitive host in Italy has already been indicated by us in Atherina boyeri.     

Record of a gregarine (Apicomplexa: Neogregarinida) in the abdomen of the termite Coptotermes gestroi (Isoptera, Rhinotermitidae)

Coptotermes gestroi is an exotic species of termite that is a pest of great economical importance in Brazil. This paper relates the occurrence of a coelomic gregarine (Apicomplexa: Neogregarinida) in the abdomen of the foraging workers recently collected from field colonies of this termite. The termite hosts presented large, white abdomens because they carried 1 up to 3 cysts of gregarines filled with numerous lemon-shaped spores. Earlier developmental stages of this gregarine were not observed in the scanning microscope preparations nor in the histological slides of the infected termites. However, the lemon-shaped spores suggest a parasite gregarine of Mattesia genus, family Lipotrophidae.     

Phylogenetic evidence for an ancestral coevolution between a major clade of coccidian parasites and elasmobranch hosts

Cartilaginous fishes are the oldest jawed vertebrates and are also reported to be the hosts of some of the most basal lineages of Cestoda and Aporocotylidae (Digenea) parasites. Recently a phylogenetic analysis of the coccidia (Apicomplexa) infecting marine vertebrates revealed that the lesser spotted dogfish harbours parasite lineages basal to Eimeria Schneider, 1875 and the group formed by Schellackia Reichenow, 1919, Lankesterella Ames, 1923, Caryospora Leger, 1904 and Isospora Schneider, 1881. In the present study we have found additional lineages of coccidian parasites infecting the cownose ray Rhinoptera bonasus Mitchill and the blue shark Prionace glauca Linnaeus. These lineages were also found as basal to species from the genera Lankesterella, Schellackia, Caryospora and Isospora infecting higher vertebrates. These results confirm previous phylogenetic assessments and suggest that these parasitic lineages first evolved in basal vertebrate hosts (i.e. Chondrichthyes), and that the more derived lineages infect higher vertebrates (e.g. birds and mammals) conforming to the evolution of their hosts. We hypothesise that elasmobranchs might host further ancestral parasite lineages harbouring unknown links of parasite evolution.     

Preliminary phylogeny of Valerianaceae (Dipsacales) inferred from nuclear and chloroplast DNA sequence data

Valerianaceae is a relatively small (ca. 350 species), but morphologically diverse angiosperm clade. Sequence data from the entire ndhF gene, the trnL-F intergenic spacer region, the trnL intron, the matK region, the rbcL-atpB intergenic spacer region and internal transcribed spacer (ITS) region of nuclear ribosomal DNA were collected for 21 taxa within Dipsacaceae and Valerianaceae (1 and 20, respectively). These data were included in several phylogenetic analyses with previously published sequences from Dipsacales. Results from these analyses (maximum parsimony, maximum likelihood, and Bayesian analysis) are in strong agreement with many of the conclusions from previous studies, most importantly: (1) Valerianaceae is sister to Dipsacaceae; (2) Triplostegia is more closely related to species of Dipsacaceae than to Valerianaceae; and (3) Valeriana appears not to be monophyletic, with Valeriana celtica falling outside the remainder of the species of Valeriana sampled here (with very strong support). With the exception of V. celtica, these data support two major clades within Valeriana; one that is exclusively New World and another that is distributed in both the Old and New World. Although the species of Valerianaceae and its sister group Dipsacaceae plus Triplostegia, are widely distributed in the Northern Hemisphere, and the data imply that Valerianaceae diversified initially in Asia (the Himalayan Patrinia and Nardostachys falling at the base of the clade), the center of modern species diversity for the group is in the Andes of South America with as many as 175 species restricted to that region. Although the exclusively South American taxa form a clade in the chloroplast and combined ITS and chloroplast analyses, support values tend to be low. Future studies will need to include additional data, in the form of both characters and taxa, before any strong conclusions about the character evolution, diversification, and biogeography of the South American valerians can be made.     

Pterospora floridiensis,a new species of acephaline gregarine (Apicomplexa) from the maldanid polychaete Axiothella mucosa in St. Andrew Bay,Florida

Pterospora floridiensis, a new species of acephaline gregarine from the body cavity of the bamboo worm Axiothella mucosa (Polychaeta: Maldanidae), is described. The gamont stage is distinctive and possesses a central cytoplasmic mass and two elongate trunks that bifurcate repeatedly and comprise approximately 60% of the total cell length. The gamont averages 198 (50–545) × 71 (25–180) m (N= 45, 43) from the tip of the trunks to the anterior (the junctional site with the other gamont in syzygy). Gametocysts average 402 (297–545) × 304 (149–495) m (N= 37). The oöcysts measure 22.5 (20.5–23.5) × 8.3 (7.0–10.0) m (N= 30) and possess an internal capsule (average length = 13.9 m, N= 30) containing the sporozoites and aliform wings on the epispore.     

Malaria parasites (Apicomplexa,Haematozoea) and their relationships with their hosts: is there an evolutionary cost for the specialization?

Parasite–host specialization is frequently considered to be a derived state such that it represents an 'evolutionary dead end' that strongly limits further evolution. In this study, it was tested whether this theory is applicable to the relationship of malaria parasites and their vertebrate hosts. For this, we revisited Perkins and Schall (2002) analysis of the phylogenetic relationships of the malaria parasites (belonging to the genera Plasmodium , Haemoproteus and Hepatocystis ) based on the mitochondrial Cytochrome b gene sequence, and inferred, using a maximum likelihood (ML) approach, the putative ancestral vertebrate hosts. As the topology in this study presents several unresolved branches and is slightly different from that of Perkins and Schall, a Shimodaira and Hasegawa (SH; 1999) test has been performed in order to properly consider several alternative topologies. The results of this study suggest that the common ancestor of all these malaria parasites was a reptile (more specific of the order Squamata), and that the host switches from Squamata to Aves and vice versa were quite frequent along the evolution of these parasites. On the contrary, a strong evidence that the host shift from Squamata to Mammalia had occurred only once during the evolution of these organisms was found. This evidence (added to the current knowledge about the association of the malaria parasites with their vertebrate hosts) allows us to suggest, at least considering the species included in this study, that the adaptation in mammals had required a high level of specialization. Hence, the acquisition of this host class had culminated in an evolutionary dead end for the mammalian malaria parasites.     

Distinct lineages of Schistocephalus parasites in threespine and ninespine stickleback hosts revealed by DNA sequence analysis

Parasitic interactions are often part of complex networks of interspecific relationships that have evolved in biological communities. Despite many years of work on the evolution of parasitism, the likelihood that sister taxa of parasites can co-evolve with their hosts to specifically infect two related lineages, even when those hosts occur sympatrically, is still unclear. Furthermore, when these specific interactions occur, the molecular and physiological basis of this specificity is still largely unknown. The presence of these specific parasitic relationships can now be tested using molecular markers such as DNA sequence variation. Here we test for specific parasitic relationships in an emerging host-parasite model, the stickleback-Schistocephalus system. Threespine and ninespine stickleback fish are intermediate hosts for Schistocephalus cestode parasites that are phenotypically very similar and have nearly identical life cycles through plankton, stickleback, and avian hosts. We analyzed over 2000 base pairs of COX1 and NADH1 mitochondrial DNA sequences in 48 Schistocephalus individuals collected from threespine and ninespine stickleback hosts from disparate geographic regions distributed across the Northern Hemisphere. Our data strongly support the presence of two distinct clades of Schistocephalus, each of which exclusively infects either threespine or ninespine stickleback. These clades most likely represent different species that diverged soon after the speciation of their stickleback hosts. In addition, genetic structuring exists among Schistocephalus taken from threespine stickleback hosts from Alaska, Oregon and Wales, although it is much less than the divergence between hosts. Our findings emphasize that biological communities may be even more complex than they first appear, and beg the question of what are the ecological, physiological, and genetic factors that maintain the specificity of the Schistocephalus parasites and their stickleback hosts.     

Ancient DNA evidence for the loss of a highly divergent brown bear clade during historical times

The genetic diversity of present-day brown bears (Ursus arctos) has been extensively studied over the years and appears to be geographically structured into five main clades. The question of the past diversity of the species has been recently addressed by ancient DNA studies that concluded to a relative genetic stability over the last 35,000 years. However, the post-last glacial maximum genetic diversity of the species still remains poorly documented, notably in the Old World. Here, we analyse Atlas brown bears, which became extinct during the Holocene period. A divergent brown bear mitochondrial DNA lineage not present in any of the previously studied modern or ancient bear samples was uncovered, suggesting that the diversity of U. arctos was larger in the past than it is now. Specifically, a significant portion (with respect to sequence divergence) of the intraspecific diversity of the brown bear was lost with the extinction of the Atlas brown bear after the Pleistocene/Holocene transition.     

Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores: new species from shrew moles (Talpidae) in the United States

All of 18 shrew moles, Neurotrichus gibbsii, collected in Oregon and Washington were infected with one or more species of coccidia. Three eimerians and one isosporan were identified and described as new species. Sporulated oocysts of Eimeria heterocapita n. sp. were subspheroid to ellipsoid, 25.5 X 21.4 (23-27 X 18-23) micron. A membranous, cap-like structure was present at one pole of the oocyst, but a micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts were 13.6 X 10.0 (12-15 X 9-11) micron; a compact sporocyst residuum was present, but Stieda, sub-, and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria neurotrichi n. sp. were ovoid, 17.6 X 13.6 (16-20 X 11-16) micron; micropyle and oocyst residuum were absent, but a polar body was present. Ovoid sporocysts were 10.7 X 5.5 (9-12 X 5-6) micron; Stieda body and sporocyst residuum were present, but sub- and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria parastiedica n. sp. were subspheroid, 27.4 X 25.5 (25-30 X 22-28) micron; micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts, pointed at both ends, were 18.3 X 10.4 (16-20 X 9-11) micron; Stieda, sub-, and parastieda bodies were present as was a sporocyst residuum. This species was found in 2 of 18 (11%) hosts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Role of gregarine parasite Ascogregarina culicis (Apicomplexa: Lecudinidae) in the maintenance of Chikungunya virus in vector mosquito

Ascogregarina culicis and Ascogregarina taiwanensis are common gregarine parasites of Aedes aegypti and Aedes albopictus mosquitoes, respectively. These mosquito species are also known to transmit dengue and Chikungunya viruses. The sporozoites of these parasites invade the midgut epithelial cells and develop intracellularly and extracellularly in the gut to complete their life cycles. The midgut is also the primary site for virus replication in the vector mosquitoes. Therefore, studies were carried out with a view to determine the possible role of these gregarines in the vertical transmission of dengue and Chikungunya viruses from larval to adult stage. Experiments were performed by exposing first instar mosquito larvae to suspensions containing parasite oocysts and viruses. Since Ascogregarina sporozoites invade the midgut of first instar larvae, the vertical transmission was determined by feeding the uninfected first instar larvae on the freshly prepared homogenates from mosquitoes, which were dually infected with viruses and the parasite oocysts. Similarly, the role of protozoan parasites in the vertical transmission of viruses was determined by exposing fresh first instar larvae to the dried pellets of homogenates prepared from the mosquitoes dually infected with viruses and the parasite oocysts. Direct vertical transmission and the vertical transmission of CHIK virus through the oocyst of the parasites were observed in the case of Ae. aegypti mosquitoes. It is suggested that As. culicis may have an important role in the maintenance of CHIK virus during the inter-epidemic period.     

DNA barcoding identifies Eimeria species and contributes to the phylogenetics of coccidian parasites (Eimeriorina, Apicomplexa, Alveolata)

Partial (～ 780 bp) mitochondrial cytochrome c oxidase subunit I (COI) and near complete nuclear 18S rDNA (～ 1,780 bp) sequences were directly compared to assess their relative usefulness as markers for species identification and phylogenetic analysis of coccidian parasites (phylum Apicomplexa). Fifteen new COI partial sequences were obtained using two pairs of new primers from rigorously characterised (sensu Reid and Long, 1979) laboratory strains of seven Eimeria spp. infecting chickens as well as three additional sequences from cloned laboratory strains of Toxoplasma gondii (ME49 and GT1) and Neospora caninum (NC1) that were used as outgroup taxa for phylogenetic analyses. Phylogenetic analyses based on COI sequences yielded robust support for the monophyly of individual Eimeria spp. infecting poultry except for the Eimeria mitis/mivati clade; however, the lack of a phenotypically characterised strain of E. mivati precludes drawing any firm conclusions regarding this observation. Unlike in the 18S rDNA-based phylogenetic reconstructions, Eimerianecatrix and Eimeria tenella formed monophyletic clades based on partial COI sequences. A species delimitation test was performed to determine the probability of making a correct identification of an unknown specimen (sequence) based on either complete 18S rDNA or partial COI sequences; in almost all cases, the partial COI sequences were more reliable as species-specific markers than complete 18S rDNA sequences. These observations demonstrate that partial COI sequences provide more synapomorphic characters at the species level than complete 18S rDNA sequences from the same taxa. We conclude that COI performs well as a marker for the identification of coccidian taxa (Eimeriorina) and will make an excellent DNA 'barcode' target for coccidia. The COI locus, in combination with an 18S rDNA sequence as an 'anchor', has sufficient phylogenetic signal to assist in the resolution of apparent paraphylies within the coccidia and likely more broadly within the Apicomplexa.     

Two malaria parasites (Apicomplexa: Plasmodiidae) of the Australian skink Egernia stokesii

The Australian skink Egernia stokesii is parasitized uncommonly by Plasmodium circularis n. sp. and by Plasmodium mackerrasae. Plasmodium circularis is distinguished from all other plasmodiids by immature schizonts that encircle host cell nuclei, forming an unbroken ring from apparent fusion of the attenuated ends. Mature schizonts contract into halteridial or dumbbell-shaped forms 15.6 x 4.3 microm, LW 66.2 microm2, with 19-52 nuclei. Rounded or oval gametocytes are 9.0 x 7.3 microm, LW 66.9 microm2, and L/W 1.24. Gametocyte LW is 2.63 x host erythrocyte nucleus size and 1.79X uninfected erythrocyte nuclei. Plasmodium mackerrasae occurs in high prevalence and often massive parasitemia in E. stokesii. Schizonts, often oblong, elongate, or oval, are 5.1 x 3.7 microm, LW 19.8 microm2, with 7.2 merozoites. Immature gametocytes, elongate with terminal nucleus, may produce multiple infections of 6 or more parasites. Mature gametocytes, usually rounded, are 5.8 x 4.6 microm, LW 26.7 microm2, and L/W 1.29. Gametocyte size is 0.98 x host erythrocyte nucleus size and 1.03 x uninfected erythrocyte nuclei. Phanerozoites, in endothelium or connective tissue of most organs, may appear in large numbers in circulating blood as seemingly intact bodies of regular form, similar to or larger than phanerozoites seen in sections. Previously unreported phenomena for hemosporidian parasites include extremely large, highly irregular exoerythrocytic schizonts, in circulating blood, perhaps torn from endothelial lining of blood vessels and sinuses, and a visible flooding of free merozoites into the blood stream.     

Longspurs and snow buntings: phylogeny and biogeography of a high-latitude clade (Calcarius)

Using complete cytochrome b sequence data, we determined that the genus Calcarius, as presently recognized, is paraphyletic. Calcarius plus Plectrophenax form a highly supported clade composed of two subclades, a "snow bunting" clade comprised of Plectrophenax plus Calcarius mccownii (formerly in the monotypic genus Rhynchophanes), and a "collared" longspur clade of Calcarius lapponicus, ornatus, and pictus. Contrary to conventional thought, Calcarius is not phylogenetically close to either Calamospiza or Emberiza. Unlike these two genera, the taxonomic affinities of Calcarius appear to lie outside of the sparrow (tribe Emberizini) assemblage. Calcarius appears to be a relatively old songbird lineage, originating between 4.2 and 6.2 million years ago. Within Calcarius, pictus and ornatus form a closely related sister pair (2.9% divergent), as do Calcarius nivalis and hyperboreus (0.18% divergent). The group (Calcarius, sensu lato) is inferred to have its origins at relatively high latitudes in the New World.     

Named species and hosts of Sarcocystis (Protozoa: Apicomplexa: Sarcocystidae)

Summary A list is given of the present 93 species of the apicomplexan protozoan genus Sarcocystis together with their definitive and intermediate hosts (if known), synonyms, homonyms, lapsi calami, etc. The names of many species of this genus are poorly known, in doubt or controversial due to lack of access to some of the literature and to failure to accept the International Code of Zoological Nomenclature.The following taxonomic innovations are introduced: New species—S. nontenella for S. tenella Eble, 1961 [non] S. tenella (Railliet, 1886) from the buzzard Buteo buteo; S. scotti for Sarcocystis sp. from the housemouse completing its sexual development in the tawny owl, Strix aluco (see Tadros & Laarman, 1980); New combinations—S. ctenosauris for Cryptosporidium ctenosauris Duszynski, 1969 from the lizard Ctenosaura similis; S. lampropeltis for Cryptosporodium lampropeltis Anderson, Duszynski & Marquardt, 1968 from the king snake Lampropeltis c. calligaster; S. roudabushi for Isospora roudabushi Pellérdy, 1974 from the gopher snake Pituophis s. sayi; and S. tropicalis for Isospora tropicalis Mukherjea & Krassner, 1965 from the golden jackal Canis aureus.Supported in part by National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland research grant AI15367.Supported in part by National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland research grant AI15367.     

Efficacy of oral sulfadimethoxine against two gregarine parasites,Protomagalhaensia granulosae and Gregarina cubensis (Apicomplexa: Eugregarinida), infecting the Death's Head cockroach,Blaberus discoidalis

Gregarines are common parasites of insects in culture, but no effective chemotherapeutic or prophylactic control protocol has been demonstrated. Sulfadimethoxine was administered in 5- and 7-day treatments to Death's Head cockroaches (Blaberus discoidalis) infected with Gregarina cubensis and Protomagalhaensia granulosae to test the efficacy of this sulfonamide against gregarine infection. Sulfadimethoxine significantly reduced the mean intensity of both G. cubensis and P. granulosae. Sulfadimethoxine treatment reduced gregarine intensity by 80% to 85% but had no significant effect on gametocyst production, suggesting that sulfonamide toxicity is directed primarily at sporozoites, trophozoites, and perhaps young gamonts. The possible use of sulfadimethoxine to produce gregarine-free insect cultures and the potential utility of gregarines as target organisms for screening pharmacologically active compounds for use against other intestinal apicomplexans are discussed.     

Phylogenetic analysis of arthropods using two nuclear protein-encoding genes supports a crustacean + hexapod clade

Recent phylogenetic analyses using molecular data suggest that hexapods are more closely related to crustaceans than to myriapods, a result that conflicts with long-held morphology-based hypotheses. Here we contribute additional information to this debate by conducting phylogenetic analyses on two nuclear protein-encoding genes, elongation factor-1 alpha (EF-1 alpha) and the largest subunit of RNA polymerase II (Pol II), from an extensive sample of arthropod taxa. Results were obtained from two data sets. One data set comprised 1092 nucleotides (364 amino acids) of EF-1 alpha and 372 nucleotides (124 amino acids) of Pol II from 30 arthropods and three lobopods. The other data set contained the same EF-1 alpha fragment and an expanded 1038-nucleotide (346-amino-acid) sample of Pol II from 17 arthropod taxa. Results from maximum-parsimony and maximum-likelihood analyses strongly supported the existence of a Crustacea + Hexapoda clade (Pancrustacea) over a Myriapoda + Hexapoda clade (Atelocerata). The apparent incompatibility between the molecule-based Pancrustacea hypothesis and morphology-based Atelocerata hypothesis is discussed.     

The complete mitochondrial DNA sequence of the crustacean Artemia franciscana

The complete mitochondrial DNA (mtDNA) sequence of the brine shrimp Artemia franciscana has been determined. It extends the present knowledge of mitochondrial genomes to the crustacean class and supplies molecular markers for future comparative studies in this large branch of the arthropod phylum. Artemia mtDNA is 15,822 nucleotides long, and when compared with its Drosophila counterpart, it shows very few gene rearrangements, merely affecting two tRNAs placed 3 downstream of the ND 2 gene. In this position a stem-loop secondary structure with characteristics similar to the vertebrate mtDNA L-strand origin of replication is found. This suggests that, associated with tRNA changes, the diversification of the mitochondrial genome from an ancestor common to crustacea and insects could be explained by errors in the mtDNA replication process. Although the gene content is the same as in most animal mtDNAs, the sizes of the protein coding genes are in some cases considerably smaller. Artemia mtDNA uses the same genetic code as found in insects, ATN and GTG are used as initiation codons, and several genes end in incomplete T or TA codons.Correspondence to: R. Garesse     

Coccidian parasites (Apicomplexa: Eimeriidae) of Graptemys caglei and Graptemys versa (Testudines: Emydidae) from Texas

Nineteen map turtles, Graptemys caglei and Graptemys versa were collected from the Guadalupe and Colorado River watersheds of south-central Texas and examined for coccidial parasites. Thirteen of the 19 turtles (68%), including 11 of 16 (69%) G. caglei and 2 of 3 (67%) G. versa, were infected with at least 1 coccidian. Five Eimeria spp. (E. chrysemydis, E. graptemydos, E. lutotestudinis, E. pseudogeographica, and E. trachemydis) were harbored by G. caglei, and 2 eimerians (E. graptemydos and E. mitraria) infected G. versa. This represents new host records for these previously described coccidians and is the first time parasites have been documented in turtles of both species.