Six new species of marine Tubificidae (Oligochaeta) from the continental shelf off Peru

Six species of marine tubifieids are described from the continental shelf off Peru. Two of them are members of the gutless genus Olavius Erséus, 1984 (subfamily Phallodrilinae). Olavius bullatus sp.n. possesses 2 pairs of large penial setae in voluminous copulatory sacs, tiny atria, and spermathecal pores in line with dorsal setae. Olavius crassitunicatus sp.n. is characterized by small atria with thin muscular layer, spermathecae with short ducts, opening dorso-laterally, and lack of penial setae. Four species arc members of the subfamily Limnodriloidinae. Limnodriloides busilicus sp.n. belongs to the appendiculatus-group (sensu Erséus). It is discriminated by somatic setae with subdental ligaments, and its voluminous elongate prostatic pads. Limnodriloides clavellatus sp.n. is distinguished from its congeners by a peculiar bulge in the cavity of each atrial ampulla, and spermatozeugmata imbedded in the walls of the spermathecae. Tectidrilus intermixtus sp.n. is similar to T. bori (Righi & Kanner, 1979); it is distinguished from the latter by having trisetal bundles in segment V or V-VI and by lacking copulatory glands. Marcusaedrilus peruanus sp.n. is characterized by nongranulated atrial ducts and bipartite spermathecae.     

Coexistence of bacterial sulfide oxidizers, sulfate reducers, and spirochetes in a gutless worm (Oligochaeta) from the Peru margin

Olavius crassitunicatus is a small symbiont-bearing worm that occurs at high abundance in oxygen-deficient sediments in the East Pacific Ocean. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization, we examined the diversity and phylogeny of bacterial symbionts in two geographically distant O. crassitunicatus populations (separated by 385 km) on the Peru margin (water depth, approximately 300 m). Five distinct bacterial phylotypes co-occurred in all specimens from both sites: two members of the gamma-Proteobacteria (Gamma 1 and 2 symbionts), two members of the delta-Proteobacteria (Delta 1 and 2 symbionts), and one spirochete. A sixth phylotype belonging to the delta-Proteobacteria (Delta 3 symbiont) was found in only one of the two host populations. Three of the O. crassitunicatus bacterial phylotypes are closely related to symbionts of other gutless oligochaete species; the Gamma 1 phylotype is closely related to sulfide-oxidizing symbionts of Olavius algarvensis, Olavius loisae, and Inanidrilus leukodermatus, the Delta 1 phylotype is closely related to sulfate-reducing symbionts of O. algarvensis, and the spirochete is closely related to spirochetal symbionts of O. loisae. In contrast, the Gamma 2 phylotype and the Delta 2 and 3 phylotypes belong to novel lineages that are not related to other bacterial symbionts. Such a phylogenetically diverse yet highly specific and stable association in which multiple bacterial phylotypes coexist within a single host has not been described previously for marine invertebrates.     

Phylogeny of 16S rRNA, ribulose 1,5-bisphosphate carboxylase/oxygenase, and adenosine 5'-phosphosulfate reductase genes from gamma- and alphaproteobacterial symbionts in gutless marine worms (oligochaeta) from Bermuda and the Bahamas

Gutless oligochaetes are small marine worms that live in obligate associations with bacterial endosymbionts. While symbionts from several host species belonging to the genus Olavius have been described, little is known of the symbionts from the host genus Inanidrilus. In this study, the diversity of bacterial endosymbionts in Inanidrilus leukodermatus from Bermuda and Inanidrilus makropetalos from the Bahamas was investigated using comparative sequence analysis of the 16S rRNA gene and fluorescence in situ hybridization. As in all other gutless oligochaetes examined to date, I. leukodermatus and I. makropetalos harbor large, oval bacteria identified as Gamma 1 symbionts. The presence of genes coding for ribulose-1,5-bisphosphate carboxylase/oxygenase form I (cbbL) and adenosine 5'-phosphosulfate reductase (aprA) supports earlier studies indicating that these symbionts are chemoautotrophic sulfur oxidizers. Alphaproteobacteria, previously identified only in the gutless oligochaete Olavius loisae from the southwest Pacific Ocean, coexist with the Gamma 1 symbionts in both I. leukodermatus and I. makropetalos, with the former harboring four and the latter two alphaproteobacterial phylotypes. The presence of these symbionts in hosts from such geographically distant oceans as the Atlantic and Pacific suggests that symbioses with alphaproteobacterial symbionts may be widespread in gutless oligochaetes. The high phylogenetic diversity of bacterial endosymbionts in two species of the genus Inanidrilus, previously known only from members of the genus Olavius, shows that the stable coexistence of multiple symbionts is a common feature in gutless oligochaetes.     

Coexistence of Bacterial Sulfide Oxidizers, Sulfate Reducers, and Spirochetes in a Gutless Worm (Oligochaeta) from the Peru Margin

Olavius crassitunicatus is a small symbiont-bearing worm that occurs at high abundance in oxygen-deficient sediments in the East Pacific Ocean. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization, we examined the diversity and phylogeny of bacterial symbionts in two geographically distant O. crassitunicatus populations (separated by 385 km) on the Peru margin (water depth, ~300 m). Five distinct bacterial phylotypes co-occurred in all specimens from both sites: two members of the γ-Proteobacteria (Gamma 1 and 2 symbionts), two members of the δ-Proteobacteria (Delta 1 and 2 symbionts), and one spirochete. A sixth phylotype belonging to the δ-Proteobacteria (Delta 3 symbiont) was found in only one of the two host populations. Three of the O. crassitunicatus bacterial phylotypes are closely related to symbionts of other gutless oligochaete species; the Gamma 1 phylotype is closely related to sulfide-oxidizing symbionts of Olavius algarvensis, Olavius loisae, and Inanidrilus leukodermatus, the Delta 1 phylotype is closely related to sulfate-reducing symbionts of O. algarvensis, and the spirochete is closely related to spirochetal symbionts of O. loisae. In contrast, the Gamma 2 phylotype and the Delta 2 and 3 phylotypes belong to novel lineages that are not related to other bacterial symbionts. Such a phylogenetically diverse yet highly specific and stable association in which multiple bacterial phylotypes coexist within a single host has not been described previously for marine invertebrates.     

A test of monophyly of the gutless Phallodrilinae (Oligochaeta, Tubificidae) and the use of a 573-bp region of the mitochondrial cytochrome oxidase I gene in analysis of annelid phylogeny

A 573-bp region of the mitochondrial gene cytochrome c oxidase subunit I (COI) of two species of Inanidrilus Erséus and four species of Olavius Erséus (Phallodrilinae, Tubificidae) is used in a parsimony analysis together with a selection of 35 other annelids (including members of Polychaeta, Pogonophora, Aphanoneura, and the clitellate taxa Tubificidae, Enchytraeidae, Naididae, Lumbriculidae, Haplotaxidae, Lumbricidae, Criodrilidae, Branchiobdellida and Hirudinea), and with two molluscs as outgroups. The data support the monophyly of the Olavius and Inanidrilus group, with a monophyletic Inanidrilus . However, parsimony jackknife analyses show that most of the other groups are unsupported by the data set, thus revealing a large amount of homoplasy in the selected gene region. Practically no information is given of within/between family relationships except for a few, closely related species. This suggests that the analysed COI region is not useful, when used alone, for inferring higher level relationships among the annelids.     

The marine Tubificidae (Oligochaeta) of the barrier reef ecosystems at Carrie Bow Cay, Belize, and other parts of the Caribbean Sea, with descriptions of twenty-seven new species and revision of Heterodrilus, Thalassodrilides and Smithsonidrilus

Fifty-nine species of Tubifieidae are recorded from Belize on the Caribbean side of Central America. Twenty-five are new to science: Ainudrilus geminus sp.n., Heronidrilus gravidus sp.n., Heterodrilus flexuosus sp.n., H. ranus sp.n., H. modestus sp.n., Coralliodrilus rugosus sp.n., C. randyi sp.n., Phallodrilus compactus sp. n., P. singularis sp.n., P. vicinus sp.n., P. nasutus sp.n., P. bipartitus sp.n., Inanidrilus reginae sp.n., Olavius finitimus sp.n., O. vacuus sp.n., O. (Olavius) pravus sp.n., Bathydrilus vetustus sp.n., B. egenus sp.n., Thalassodrilides bruneti sp.n., Limnodriloides anxius sp.n., L. major sp.n., L. sacculus sp.n., L. adversus sp.n., Smithsonidrilus appositus sp.n., S. involutus sp.n. Phylogenetic analyses indicate that Heterodrilus Pierantoni, 1902, Thalassodrilides Brinkhurst & Baker, 1979, and Marcusaedrilus Righi & Kanner, 1979, are paraphyletic taxa as previously defined. They are therefore revised to include also apomorphic species earlier attributed to separate genera; in so doing, Marcusaedrilus becomes a junior synonym of Smithsonidrilus Brinkhurst, 1966. Smithsonidrilus pauper sp.n. (Peru), and S. multiglandularis sp.n. (Florida and Puerto Rico) are also described, and Limnodriloides claviger Erséus, 1982 is regarded as a synonym of L. pierantonii (Hrabě, 1971).     

Cuticular ultrastructure in some marine oligochaetes (Tubificidae)

Abstract. The ultrastructure of the thin, non-cellular cuticle is described for 6 marine oligochaetes, representing 3 of the subfamilies (Phallodrilinae, Limnodriloidinae, and Rhyacodrilinae) of the Tubificidae. The main components of the cuticle in these 6 species, as in most other oligochaetes examined, are: (1) a fiber zone closest to the epidermis, consisting of collagen fibers embedded in a matrix, (2) an epicuticle, which is a continuation of the matrix outside the fiber zone, and (3) epicuticular projections, which are membrane-bound bodies covering the outer surface of the epicuticle. The projections are probably formed by the microvilli that penetrate the cuticle from the epidermal cells below, but this was confirmed only in the studied limnodriloidines. Three of the species examined, Duridrilus turdus, Olavius vacuus , and Heterodrilus paucifascis , lack microvilli. The morphology of the components in the cuticle differs between the studied species. The collagen fibers may form an "orthogonal grid" (i.e., layers of parallel fibers perpendicular to the layers immediately above and below), or they may form parallel layers, or be irregularly scattered. The number of dense layers in the epicuticle, as well as the shape and internal structure of the epicuticular projections, also vary. All these characters might be useful in future phylogenetic analyses to achieve better hypotheses of relationships within oligochaetes as well as to other groups.     

A triple bacterial endosymbiosis in a gutless oligochaete (Annelida): ultrastructural and immunocytochemical evidence

Abstract. The gutless marine oligochaete, Olavius crassitunicatus finogenova 1986 (Tubificidae), from suboxic to sulfidic sediments off Peru, consistently harbored 3 structurally distinct types of extracellular bacterial symbionts. Large, oval bacteria were labeled immunocytochemically, proving their nature as autotrophs. Spectroscopical analysis documented storage of sulfur in this numerically dominant morphotype. Small, rod-shaped bacteria attained a more peripheral position adjacent to the cuticle. The third bacterial type was represented by long, filamentous forms which were often in close contact to the oval bacteria. With their curved or undulate cells, these filiform bacteria resembled spirochetes. They were clearly distinguishable and consistently found in all investigated host specimens.
While molecular analyses could not be performed, structural and immunocytochemical evidence indicated that the oval bacteria seemed equivalent to the γ-proteobacteria in related gutless oligochaetes. On the basis of morphological similarity and indications from closely related symbiotic tubificids, the possible relationship of the two other morphoptypes must remain unsolved and needs further molecular analysis. The three bacterial morphotypes live in a consistent, elaborate, and apparently obligate coexistence with a host that has completely reduced its digestive and excretory organs.     

Phylogeny of 16S rRNA, Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase, and Adenosine 5′-Phosphosulfate Reductase Genes from Gamma- and Alphaproteobacterial Symbionts in Gutless Marine Worms (Oligochaeta) from Bermuda and the Bahamas

Gutless oligochaetes are small marine worms that live in obligate associations with bacterial endosymbionts. While symbionts from several host species belonging to the genus Olavius have been described, little is known of the symbionts from the host genus Inanidrilus. In this study, the diversity of bacterial endosymbionts in Inanidrilus leukodermatus from Bermuda and Inanidrilus makropetalos from the Bahamas was investigated using comparative sequence analysis of the 16S rRNA gene and fluorescence in situ hybridization. As in all other gutless oligochaetes examined to date, I. leukodermatus and I. makropetalos harbor large, oval bacteria identified as Gamma 1 symbionts. The presence of genes coding for ribulose-1,5-bisphosphate carboxylase/oxygenase form I (cbbL) and adenosine 5′-phosphosulfate reductase (aprA) supports earlier studies indicating that these symbionts are chemoautotrophic sulfur oxidizers. Alphaproteobacteria, previously identified only in the gutless oligochaete Olavius loisae from the southwest Pacific Ocean, coexist with the Gamma 1 symbionts in both I. leukodermatus and I. makropetalos, with the former harboring four and the latter two alphaproteobacterial phylotypes. The presence of these symbionts in hosts from such geographically distant oceans as the Atlantic and Pacific suggests that symbioses with alphaproteobacterial symbionts may be widespread in gutless oligochaetes. The high phylogenetic diversity of bacterial endosymbionts in two species of the genus Inanidrilus, previously known only from members of the genus Olavius, shows that the stable coexistence of multiple symbionts is a common feature in gutless oligochaetes.     

Lindernia kinmenensis sp. nov. (Scrophulariaceae) from Kinmen (Taiwan)

Lindernia kinmenensis Y. S. Liang, C. H. Chen, & C. L. Tasi sp. nov. from Kinmen (Taiwan) is described. The new species belongs to section Torenioides and is most similar to L. crustacea. It differs from the latter by the following characters: calyx pubescent between the ridges (vs pubescent on ridges), with (2)3(4) bristles on the ridge near the apex of the calyx‐lobes (vs without such bristles), corolla shorter, 6–8 mm long (vs 7–11 mm long) and pale purple (vs purple or blue), posterior theca with obtuse (vs acute) apex. This species is usually found in sandy wetlands. A line drawing, colour photos and SEM micrographs of pollen and seed as aids for identification are provided.     

Heliconema minnanensis n. sp. (Physalopteroidae: Physalopteridae) and Raphidascaris trichiuri (Yin and Zhang) n. comb. (Ascaridoidea: Anisakidae) in marine fishes.

Heliconema minnanensis n. sp. is described from the Chiloscyllium plagiosum south of the Minnan-Taiwan Bank Fishing Ground, Taiwan Strait. It is characterized by having the following combination of features. There are no longitudinal ridges in the posterior part of the body; the male has 4 pairs of precloacal papillae and 6-7 pairs of postcloacal papillae. The right spicule averages 0.2 mm long (1.1% of body length), and the left spicule averages 1.2 mm long (5.6% of body length), making an average spicule ratio of 1:5.2. The female has a vulva located on the left side of a thick membrane and 56-65% of the body length from the anterior end. Raphidascaris trichiuri (Yin and Zhang, 1983) comb. n., known previously from males only, is transferred from Cloeoascaris and redescribed, including material of both sexes from Muraenesox cinereus from the type locality.     

ONSLOWIACEAE FAM. NOV. (PHAEOPHYCEAE)

We propose the separation of Onslowia and Verosphacella from the Choristocarpaceae and from the Sphacelariales based on comparisons of DNA sequences of rbc L and nrRNA genes and morphological considerations. The new family Onslowiaceae is created to include these two genera. The families Choristocarpaceae and Onslowiaceae are considered incertae sedis.     

Rhabditis (Oscheius) pheropsophi n. sp. (Rhabditida: Rhabditidae)

Rhabditis (Oscheius) pheropsophi n. sp., associated with cadavers of the bombardier beetle, Pheropsophus aequinoctialis, is described from material collected in Brazil. Mean body length of the female is 1,217 μm, of the male 872 μm, and of the dauer juvenile 568 μm. The female has six lips with one papilla on each lateral lip and two on each sublateral lip; stoma wall thickened dorsally, metarhabdions with warts, excretory pore near base of esophagus, tail long (c = 8), and phasmids prominent, protruding on scanning electron microscope preparations. The male has 10 pairs of bursal ribs, with the terminal pair considerably smaller than the others; spicules fused distally two-thirds of their length. The new species can be distinguished from other members of the Dolichura-group by its fused spicules.     

High content of proteins containing 21st and 22nd amino acids, selenocysteine and pyrrolysine, in a symbiotic deltaproteobacterium of gutless worm Olavius algarvensis

Selenocysteine (Sec) and pyrrolysine (Pyl) are rare amino acids that are cotranslationally inserted into proteins and known as the 21st and 22nd amino acids in the genetic code. Sec and Pyl are encoded by UGA and UAG codons, respectively, which normally serve as stop signals. Herein, we report on unusually large selenoproteomes and pyrroproteomes in a symbiont metagenomic dataset of a marine gutless worm, Olavius algarvensis. We identified 99 selenoprotein genes that clustered into 30 families, including 17 new selenoprotein genes that belong to six families. In addition, several Pyl-containing proteins were identified in this dataset. Most selenoproteins and Pyl-containing proteins were present in a single deltaproteobacterium, δ1 symbiont, which contained the largest number of both selenoproteins and Pyl-containing proteins of any organism reported to date. Our data contrast with the previous observations that symbionts and host-associated bacteria either lose Sec utilization or possess a limited number of selenoproteins, and suggest that the environment in the gutless worm promotes Sec and Pyl utilization. Anaerobic conditions and consistent selenium supply might be the factors that support the use of amino acids that extend the genetic code.     

Cryptosporidium suis n. sp. (Apicomplexa: Cryptosporidiidae) in pigs (Sus scrofa)

Molecular and biological characteristics of a new species of Cryptosporidium from the feces of pigs (Sus scrofa) is described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum; they are passed fully sporulated, lack sporocysts, and measure 4.9-4.4 microm (mean = 4.6 microm) x 4.0-4.3 microm (mean = 4.2 microm); length to width ratio 1.1 (n = 50). Cryptosporidium suis is not transmissible to nude mice and is poorly infectious for cattle. Molecular and phylogenetic analyses at the 18S ribosomal RNA, heat shock protein 70, and actin gene loci demonstrate C. suis to be genetically distinct from all known species and genotypes of Cryptosporidium, and thus is named as Cryptosporidium suis.     

Redescription of Monhystrella parvella (Filipjev) comb.n. (Nematoda, Monhysteridae)

The type of material of Monhystrella parvella (Filipjev, 1931) comb.n. from Abyssinia (Ethiopia) is redescribed and figured in detail. Data on the genital system and the ontogeny of this species are given. Monhystera filiformis sensu Gerlach, 1951 nec Bastian, 1865 of the Black Sea at Pomorie (Bulgaria) is synonymised with M. parvella . The Black Sea specimens have a shorter pharynx, a longer tail and differ also from the Abyssinian specimens in the shape of the tail and the spinneret outlet.     

Megalophallus deblocki n. sp. (Digenea: Microphallidae) from Rostrhamus sociabilis (Vieillot) (Aves: Accipitridae) in Paraguay

A new species in the little-known digenean fauna of Paraguayan birds is described in detail, especially in relation to its terminal genitalia. The new form, Megalophallus deblocki n. sp., is a microphallid from the intestine of the snail kite Rostrhamus sociabilis (Vieillot). It is differentiated in detail from its congeners, but differs mainly in having a substantially larger phallus (e.g. in relation to the ventral sucker), a smaller ventral sucker and smaller eggs. Comments are included on the functional morphology of the terminal genitalia.     

Carnoya kermarreci n.sp. (Rhigonematidae: Nematoda) from Anadenobolus politus (Porat) (Rhinocricidae: Diplopoda) from Guadeloupe

Summary Carnoya kermarreci n.sp. (Rhigonematidae; Nematoda) is described from Anadenobolus politus (Porat) (Rhinocricidae; Diplopoda) from Guadeloupe. The new species most closely resembles C. martiniquensis Adamson, 1984 from A. politus in Martinique and C. venezuelensis Adamson, 1984 from Rhinocricus flavocinctus from Venezuela in that the lips in the male are broad and the arrangement of caudal papillae is almost identical in the three species. Males of C. kermarreci are easily distinguished by the absence of spines, by the form of the cuticular projections at the base of the buccal cavity and by the fact that the most anterior pair of caudal papillae are located very close together. Females are distinguished by the presence of three pairs of somatic papillae near the level of the vulva. The nematode fauna of A. politus includes species from two distinct orders and presumably reflects the fact that diplopods are among the most ancient hosts of nematodes.     

Rhabditis (Rhabditis) freitasi sp.n. and Rhabditis (Rhabditis) costai sp.n. (Nematoda-Rhabditidae) isolated from bovine otitis

The author described two new species of Nematoda-Rhabditidae collected from the auditory meatus of catle with ear infection: Rhabditis (Rhabditis) freitasi sp.n. from Formosa county in the state of Goiás and Rhabditis (Rhabditis) costai sp.n. from Sert?ozinho county in the state of S?o Paulo.