(25S)-Cholesten-26-oic acid derivatives from an Indonesian soft coral Minabea sp.

(25S)-3-Oxocholesta-1,4-dien-26-oic acid (1) and a new (25S)-18-acetoxy-3-oxocholesta-1,4-dien-26-oic acid (2) were isolated from a soft coral Minabea sp. (cf. aldersladei) collected in North Sulawesi, Indonesia, together with two known cholic-acid-type compounds, 3-oxochol-1,4-dien-24-oic acid (3) and 3-oxochol-4-en-24-oic acid (4). The structures of these compounds were determined on the basis of their spectroscopic data. The absolute stereochemistry at C-25 of 2 was determined by comparative ¹H NMR study using chiral anisotropic reagents [(S)- and (R)-phenylglycine methyl esters]. This is the first to report compound 1 as a natural product.     

Eimeria clethrionomyis sp. n., Eimeria gallatii sp. n., Eimeria pileata sp. n. and Eimeria marconii sp. n. from the Red-Backed Vole Clethrionomys gapperi Vigors,from Pennsylvania§

SYNOPSIS Four new eimerian species are described from red-backed voles. Clethrionomys gapperi in Pennsylvania. Sporulated oocysts of Eimeria clethrionomyis sp. n. are ellipsoidal, 18.8 (16.5–21.5) × 14.9 (14.0–16.5) with elongate, ovoid sporocysts, 10.6 (9.5–12.0) × 6.1 (5.5–7.0). The oocyst wall is smooth, with 2 layers, and thins, with terminal cap at one or both ends. Polar granules, dark Stieda body and sporocyst residuum are present. The occyst residuum is absent. Sporulated oocysts of Eimeria gallatii sp. n. are ellipsoidal, 27.7 (21–32) × 19.3 (17–24) with ovoid sporocysts, 13.5 (12–15) × 8.8 (8–10). The oocyst wall is smooth, 2-layered, with a micropyle and thin wall at the end opposite the micropyle. Polar granules. Stieda body and sporocyst residuum are present. The oocyst residuum is atypical, of cobwebby material. Sporulated oocysts of Eimeria pileata sp. n. are subspherical to spherical, 25.2 (20.5–29.5) × 22.5(19.5–25.5) with ellipsoidal sporocysts, 13.4(10.5–15.0) × 8.4 (7.5–9.5). The oocyst wall is rough, pitted, striated, 2-layered, with no micropyle. Polar granules, oocyst and sporocyst residuum. Stieda body and stiedal cap are present. Sporulated oocysts of Eimeria marconii sp. n. are ellipsoidal, 13.0 (10.5–15.0) × 10.6 (9.5–12.0) with elongate, ovoid sporocysts, 7.7 (7.0–8.5) × 4.2 (3.0–4.5). The oocyst wall is smooth, single-layered, with no micropyle. Polar granules, dark Stieda body and sporocyst residuum are present. There is no oocyst residuum.     

Longidorus grandis n. sp. and L. paralongicaudatus n. sp. (Nematoda: Longidoridae), Two Parthenogenetic Species from Arkansas

Two new parthenogenetic species of Longidorus were found in Arkansas. Longidorus grandis n. sp. is characterized by its body (5.80-8.24 mm), slightly offset head, head width 20-27 µm, odontostyle 86-100 µm, guide ring 26-35 µm posterior to the anterior end, short conoid to mammiliform tail. Longidorus grandis n. sp. is similar to L. vineacola Sturhan &Weischer, 1964; L. lusitanicus Macara, 1985; L. edmundsi Hunt &Siddiqi, 1977; L. kuiperi Brinkman, Loof &Barbez, 1987; L. balticus Brzeski, Peneva &Brown, 2000; L. closelongatus Stoyanov, 1964; and L. seinhorsti Peneva, Loof &Brown, 1998. Longidorus paralongicaudatus n. sp. is characterized by its body length (2.60-5.00 µm), anteriorly flattened and offset head region 13-18 µm wide, odontostyle length 92-127 µm, guide ring 21-30 µm posterior to the anterior end, tail elongate-conical, and c'' = 1.2-2.6. Longidorus paralongicaudatus n. sp. most closely resembles L. longicaudatus Siddiqi, 1962; L. socialis Singh &Khan, 1996; L. juvenilis Dalmasso, 1969; and L. curvatus Khan, 1986.     

Descriptions of Deladenus albizicus n. sp. and D. processus n. sp. (Nematoda: Hexatylina) from Haryana,India

Two different nematodes were isolated from the bark of Albizia lebbeck trees; one from insect infested and another from noninfested, healthy tree. Based on the biological, morphological, and molecular evidences, the nematodes are described as Deladenus albizicus n. sp. and D. processus n. sp. (Nematoda: Hexatylina). Deladenus albizicus n. sp., isolated from insect-infested tree, multiplied on the fungus Nigrospora oryzae. Myceliophagous females of this nematode reproduced by parthenogenesis and spermathecae were indistinct. Infective females, readily produced in the cultures, are dorsally curved. Only one type of males containing small-sized sperms in their genital tracts were produced in the culture. Myceliophagous females: L = 0.75 to 1.71 mm, a = 32.3 to 50.8, b = 9.3 to 11.2, b’ = 5.2 to 7.3, c = 27.2 to 35.6, V = 91.0 to 93.3, c’ = 2.0 to 2.9, stylet = 11 to 12 µm, excretory pore in the region of median pharyngeal bulb, 43 to 47 µm anterior to hemizonid. Deladenus processus n. sp., isolated from bark of healthy A. lebbeck tree, was cultured on Alternaria alternata. Myceliophagous females reproduced by amphimixis and their spermathecae contained rounded sperms. Infective females were never produced, even in old cultures. Myceliophagous females: L = 0.76 to 0.99 mm, a = 34 to 49, b = 13.3 to 17.7, b’ = 3.8 to 5.8, c = 19.6 to 22.8, V = 92.2 to 93.5, c’ = 2.7 to 3.5, stylet = 6 to 7 µm, excretory pore in the proximity of hemizonid, tail conoid, tapering from both sides to a long pointed central process. It is proposed to classify Deladenus species in three groups: durus, siricidicola, and laricis groups based on female and spermatogonia dimorphism, mode of reproduction, and insect parasitism.     

Halorubellus salinus gen. nov., sp. nov. and Halorubellus litoreus sp. nov., novel halophilic archaea isolated from a marine solar saltern

Two extremely halophilic archaeal strains GX3(T) and GX26(T) were isolated from the Gangxi marine solar saltern near the Weihai city of Shandong Province, China. Cells from the two strains were pleomorphic and stained Gram-negative, colonies were red-pigmented. Strains GX3(T) and GX26(T) were able to grow at 25-50 °C (optimum 37 °C), at 1.4-5.1M NaCl (optimum 3.1M), at pH 5.5-9.5 (optimum pH 7.0) and neither strain required Mg(2+) for growth. Cells lyse in distilled water and the minimal NaCl concentration to prevent cell-lysis was 8% (w/v). The major polar lipids of the two strains were PA (phosphatidic acid), PG (phosphatidylglycerol), PGP-Me (phosphatidylglycerol phosphate methyl ester) and three major glycolipids (GL1, GL2 & GL3) chromatographically identical to S-TGD-1 (sulfated galactosyl mannosy glucosyl diether), S-DGD-1 (sulfated mannosyl glucosyl diether), and DGD-1 (mannosyl glucosyl diether) respectively, an unidentified lipid (GL4) was also detected in strain GX26(T). Phylogenetic analysis based on 16S rRNA gene revealed that strain GX3(T) and strain GX26(T) formed a distinct clade with the closest relative, Haladaptatus paucihalophilus (89.9-92.4% and 90.4-92.7, respectively). The rpoB' gene similarities between strains GX3(T) and GX26(T), and between the two strains and the closest relative, Halorussus rarus TBN4(T) are 96.5%, 84.3% and 83.9%, respectively. The DNA G+C contents of strain GX3(T) and strain GX26(T) are 67.3 mol% and 67.2 mol%, respectively. The DNA-DNA hybridization value between strain GX3(T) and strain GX26(T) was 44%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX3(T) and strain GX26(T) represent two novel species in a new genus within the family Halobacteriaceae, Halorubellus salinus gen. nov., sp. nov. (type strain GX3(T)=CGMCC 1.10384(T)=JCM 17115(T)) and Halorubellus litoreus sp. nov. (type strain GX26(T)=CGMCC 1.10386(T)=JCM 17117(T)).     

Cryptosporidium molnari n. sp. (Apicomplexa: Cryptosporidiidae) infecting two marine fish species,Sparus aurata L. and Dicentrarchus labrax L

Cryptosporidium molnari n. sp. is described from two teleost fish, the gilthead sea bream (Sparus aurata L.) and the European sea bass (Dicentrarchus labrax L.). The parasite was found mainly in the stomach epithelium and seldom in the intestine. Oocysts were almost spherical, with four naked sporozoites and a prominent residuum, and measured 3.23-5.45 x 3.02-5.04 (mean 4.72 x 4.47) microm in the type host, gilthead sea bream (shape index 1-1.17, mean 1.05). Sporulation was endogenous, as fully sporulated oocysts were found within the fish, both in the stomach epithelium and lumen, and in faeces. Oocysts and other stages of C. molnari fit most of the diagnostic features of the genus Cryptosporidium, but differ from hitherto described species, including piscine ones. All stages were located within a host contributed parasitophorous vacuole lined by a double host microvillar membrane. Merogonial and gamogonial stages appeared in the typical extracytoplasmic position, whereas oogonial and sporogonial stages were located deeply within the epithelium. Ultrastructural features, including the characteristic contact zone of the parasite with the host epithelial surface, were mostly coincident with those of other Cryptosporidium spp. Mitochondria were found in dividing meronts, merozoites, microgamonts and sporozoites. Pathological effects were more evident in gilthead sea bream, which also exhibited a clearly higher prevalence (24.4 versus 4.64% in sea bass). External clinical signs, consisting of whitish faeces, abdominal swelling and ascites, were rarely observed, in contrast with important histopathological damage. The wide zones of epithelium invaded by oogonial and sporogonial stages appeared necrotic, with abundant cell debris, and sloughing of epithelial cells, which detached to the lumen. No inflammation reaction was observed and the cellular reaction was limited to the cells involved in the engulfing of intraepithelial stages and debris, probably macrophages.     

Haemogregarina pseudemydis n. sp. (Apicomplexa: Haemogregarinidae) and Pirhemocyton chelonarum n. sp. in Turtles from Louisiana*

SYNOPSIS. The blood of each of 95 turtles (8 species) collected from southeastern Louisiana was infected with some or all of the merogonic stages and gametocyte stage of Haemogregarina pseudemydis n. sp. Five species of turtles harbored Pirhemocyton chelonarum n. sp. Turtle Haemogregarina and Pirhemocyton are locality records for Louisiana. Pirhemocyton is reported for the first time in turtles and in the continental U.S.A.     

Tasmanomysis oculata n.g. n.sp. from Tasmanian waters (Crustacea: Mysidacea: Mysini)

Tasmanomysis oculata n.g. n.sp. belonging to the Tribe Mysini is described from the coastal waters of southern Tasmania. This new genus superficially resembles the monospecific genus Arthromysis Colosi,1924 known only from the Straits of Magellan. The form of the pleopods in the male together with the segmentation of the thoracic legs and shape of the telson clearly distinguish the two genera.     

Steinernema aciari sp. n. (Nematoda: Steinernematidae), a new entomopathogenic nematode from Guangdong, China

A new species of entomopathogenic nematode, Steinernema aciari sp. n. was described. It was recovered from a soil sample collected from Haimen town, Shantou district in the eastern coast of Guangdong province, the People's Republic of China during a survey for entomopathogenic nematodes. S. aciari sp. n. belongs to the Steinernema glaseri group. It can be separated from all described Steinernema species by the combined morphological and morphometrical characters of various stages of the nematodes. For male, the new species can be recognized by spicule length (86+/-6.3 microm); spicule tip blunt with a hook-like structure; gubernaculum with a short and Y-shaped cuneus and corpus well-separated posteriorly. For infective juvenile, the combination of the following characters: body length (1113+/-68 microm), distance from anterior end to excretory pore (95+/-3.7 microm), tail length (78+/-5.2 microm), and E % (123+/-7) can be used to differentiate the new species from other nematodes. For female, the tail (conoid with a long mamillate terminus and a distinct postanal swelling) and vulva (slightly protruding from body surface with conspicuous double flapped epiptygma) shapes can be used as diagnostic characters for the new species. The new species can also be distinguished from other Steinernema species by DNA sequences of either a partial 28S rDNA or the internal transcribed spacer regions of rDNA, and from the close related species S. glaseri, Steinernema longicaudum CWL05, and Steinernema guangdongense by cross-breeding test.     

Phyllodiaptomus praedictus n. sp. (Copepoda,Calanoida) from Thailand

Phyllodiaptomus praedictus n. sp. is described from the Bangkok area. It might be endemic to Thailand. It belongs to the blanci-group of species, and is most closely related to P. annae and P. wellekensae.     

A new microsporidian species, Vairimorpha ocinarae n. sp., isolated from Ocinara lida Moore (Lepidoptera: Bombycidae) in Taiwan

A new microsporidium was isolated from Ocinara lida Moore (Lepidoptera: Bombycidae), a pest of Ficus microcarpa L. f. in Taiwan. The microsporidium produces systemic infections in O. lida larvae; the midgut epithelium, Malpighian tubules, and midgut muscle tissues were the target tissues for this isolate, and atrophied fat body tissues were found in heavily infected larvae. Two types of spores were observed, diplokaroytic spores with 11-13 coils of polar tube, and monokaryotic spores with 12 coils of the polar tube that developed within a sporophorous vesicle to form octospores. Electron-dense granules were abundant in the episporontal space of the sporophorous vesicles, and were similar to those of Vairimorpha invictae isolated from Solenopsis invicta, but different from granules or inclusions of other Vairimorpha species. Based on the phylogenetic analysis of the small subunit ribosomal DNA sequence, this isolate is unique within the Vairimorpha complex. Morphological and genetic characters showed this isolate to be a new species. It is placed in the genus Vairimorpha and is described as Vairimorpha ocinarae n. sp.     

Eodiaptomus shihi n. sp. (Copepoda,Calanoida) from central India

Eodiaptomus shihi n. sp. is described from the material collected from Gandhisagar reservoir, River Narmada and a hillpool in the Madhya Pradesh State of central India. The salient features of this new species include: in female, the left metasomal wing is strongly developed, and the endopodite in leg 5 is 2-segmented and longer than first exopodite-segment; in male, the right caudal ramus is armed with a tooth-like chitinous structure near ventro-distal region, and in right leg 5, the basis is devoid of any hyaline lobe on inner margin and the first exopodite-segment is produced into a large spinous process at disto-outer corner.     

Facieplatycauda pratti gen. n., sp. n. and Two New Species of Myxobolus (Myxosporida)*

Synopsis. Facieplatycauda pratti gen. n., sp. n. and Myxobolus kozloffi sp. n., from the Klamath Lake sucker, Catostomus luxatus (Cope), are described. The new genus is placed in the family Myxobolidae between Henneguya and Myxobolus. The spore valves beyond the sporoplasmic space are broad and long but not attenuated as an inverted triangle as in certain species of Myxobolus with tails, or of Henneguya with sometimes bifurcated tails. An organism resembling Myxobolus insidiosus (Wyatt & Pratt) was found in the muscle of the body of Salmo clarki (Richardson). Differences in spore structure and site of infection suggest this is a subspecies of the previous form which should be designated Myxobolus insidiosus clarki. A previously unreported species of Myxobolus from Cottus aleuticus (Gilbert) is also described.     

Eimeria cicaki sp. n. and Isospora thavari sp. n. from the House Lizard Gehyra mutilata Boulenger in Malaysia*

SYNOPSIS. Oocysts and endogenous stages of new species of Eimeria and Isospora from the house lizard, Gehyra mutilata, are described. The ellipsoid to subspherical 2-layered oocysts of E. cicaki averaged 24.0 × 21.0 μm. Polar granules are present. Micropyle and oocyst residuum are absent. Ellipsoid sporocysts average 12.2 × 9.0 μm. A sporocyst residuum is present, but the Stieda body is absent. Endogenous stages are in epithelial cells of the small intestine. The subspherical single-layered oocysts of I. thavari average 23.8 × 22.8 μm. The polar granule is present; micropyle and oocyst residuum are absent. Ellipsoid sporocysts average 12.8 × 9.4 μm. Stieda body and sporocyst residuum are present. There are endogenous stages in epithelial cells of the small intestine.     

Isolation and absolute stereochemistry of coussaric acid,a new bioactive triterpenoid from the stems of Coussarea brevicaulis

Coussaric acid (1), a triterpenoid based on an ursane skeleton, and an oleanane-type triterpene acid, 3-epi-spathodic acid (2), as well as four known compounds, barbinervic acid, scutellaric acid, stigmasterol and stigmasterol glucoside, have been isolated from an EtOAc-soluble extract of the stems of Coussarea brevicaulis. The structures of compounds 1 and 2 were elucidated on the basis of spectroscopic investigation, and single-crystal X-ray crystallography was used to confirm the structure of 1. The absolute stereochemistry of 1 was established by chemical transformations and by the Mosher ester procedure. The potential of the isolates and chemical transformation products to induce quinone reductase was evaluated in mouse Hepa lclc7 hepatoma cells.     

Prolagus italicus n. sp. (Ochotonidae, Lagomorpha), a new Pliocene species of peninsular Italy

Prolagus italicus n. sp., a new endemic Pliocene species of peninsular Italy, is described here, adding new details to the knowledge of this widespread and long-lasting ochotonid genus. Several dental morphodimensional characters suggest a close relationship between P. italicus and P. sorbinii (evaporitic and postevaporitic Messinian, central Italy). On the other hand, P. italicus shares with other Plio-Pleistocene Prolagus species of continental Europe a large size and some peculiar characters of dental occlusal surface. Actually, an evolutionary trend can be detected in Plio-Pleistocene species of the genus Prolagus. The value of P. italicus in biochronology is quite consistent, as this species seems to mark MN17 in central Italy. Moreover, some morphodimensional differences between P. italicus populations have been identified: this can help to infer relative chronological positions to the fossil sites in which P. italicus is present. Some morphological characters present in one of the two available P. italicus populations are apparently related to ecological more than to evolutionary factors: this may be a starting point for in-depth palaeoecological studies related to the genus Prolagus. The presence of P. italicus in the Italian peninsula may have prevented the entrance of other species of western European affinity.     

Photobacterium piscicola sp. nov., isolated from marine fish and spoiled packed cod

Five isolates from marine fish (W3^T, WM, W1S, S2 and S3) and three isolates misclassified as Photobacterium phosphoreum, originating from spoiled modified atmosphere packed stored cod (NCIMB 13482 and NCIMB 13483) and the intestine of skate (NCIMB 192), were subjected to a polyphasic taxonomic study. Phylogenetic analysis of 16S rRNA gene sequences showed that the isolates were members of the genus Photobacterium. Sequence analysis using the gapA, gyrB, pyrH, recA and rpoA loci showed that these isolates formed a distinct branch in the genus Photobacterium, and were most closely related to Photobacterium aquimaris, Photobacterium kishitanii, Photobacterium phosphoreum and Photobacterium iliopiscarium. The luxA gene was present in isolates W3^T, WM, W1S, S2 and S3 but not in NCIMB 13482, NCIMB 13483 and NCIMB 192. AFLP and (GTG)₅-PCR fingerprinting indicated that the eight isolates represented at least five distinct genotypes. DNA–DNA hybridizations revealed 89% relatedness between isolate W3^T and NCIMB 192, and values below 70% with the type strains of the phylogenetically closest species, P. iliopiscarium LMG 19543^T, P. kishitanii LMG 23890^T, P. aquimaris LMG 26951^T and P. phosphoreum LMG4233^T. The strains of this new taxon could also be distinguished from the latter species by phenotypic characteristics. Therefore, we propose to classify this new taxon as Photobacterium piscicola sp. nov., with W3^T (=NCCB 100098^T = LMG 27681^T) as the type strain.     

Senecella siberica n. sp. and the position of the genus Senecella in Calanoida classification

Senecella calanoides Juday has been reported from fresh-water lakes of North America, Northern Siberia and inshore brackish waters of Kara and Laptev Seas. Investigation of 11 females and 2 males from the brackish waters of Neelov Bay (Laptev Sea) shows that second species, Senecella siberica sp. n., can be distinguished from S. Calanoides Juday by the greater size, a slightly asymmetrical female genital segment, the 3 external spines on the 3d exopod of 2 to 4th swimming legs and by more longer left endopod digitiform process of the male 5th legs. S. siberica seems to be brackish-water euryhaline species, inhabiting brackish waters with salinity from 1 to 22. The genus belongs to the family Aetideidae and is the only genus with freshwater representatives.     

Mongolodiaptomus dumonti n. sp., a new freshwater copepod (Calanoida,Diaptomidae) from Thailand

Mongolodiaptomus dumonti n. sp. from several localities in northeast Thailand is described and figured. The new species shares some features with Mongolodiaptomus gladiolus (Shen & Lee), M. calcarus (Shen & Tai) and M. rarus (Reddy, Sanoamuang & Dumont), thus the gladiolus-group is created. However, it stands out in the female, by the extraordinary large postero-laterally spine at left proximal margin of genital somite. In the male, the right caudal ramus has a large tooth and a bilobed knob on ventral side; basis of the right P5 has a distinct, spur-like process on mid-distal surface; the second exopodite-segment of the right P5 has characteristically irregular outer margin, enlarged at proximal 1/3, but narrowed at distal 2/3 and bearing three lateral spines of unequal sizes; the inner margin of basis of the left P5 is fringed with a narrow hyaline lamella and dilated distally in lateral angle.