<Emphasis Type="Italic">Epigonus cavaticus</Emphasis> (Teleostei: Perciformes), a new epigonid fish from Palau,western Central Pacific

A new epigonid fish, Epigonus cavaticus, is described on the basis of eight specimens (59.2–69.5 in standard length: SL) collected from a cave at depth 20 m, southern fringing reef of Ngemelis Island, Palau. The species differs from other congeners by having minute teeth on both jaws, no opercular spine, pyloric caeca 7–8, gill rakers 25–27, total pored lateral line scales 48–50, dorsal fin rays VII-I, 10–11 (mode VII-I, 10), pectoral fin rays 16, vertebrae 10 + 15, body depth 21.4–25.0% SL, pectoral fin length 22.7–24.6% SL, eye diameter 44.4–47.5% head length: HL, upper jaw length 40.2–42.5% HL. Four paratypes (63.7–66.8 mm SL) of the new species are female with mature gonads, it is the smallest in size at sexual maturity among the congeners.     

Two new and two resurrected species of the sciaenid genusJohnius (Johnius) from the West Pacific

Two new species of the sciaenid genusJohnius (Johnius) are described:J. trewavasae from Taiwan, Hong Kong, and Singapore, differs from all other congeners in having 24–27 dorsal soft rays, 5–6 scales above and 7–10 scales below the lateral line, 6–8 obtuse lower gill rakers, the last pleural rib on the 11th vertebra, and a shorter lower jaw (33.8–38.4% HL);J. latifrons from Thailand and Java is characterized by 25–29 dorsal soft rays, 7–9 scales above and 11–14 scales below the lateral line, 7–9 obtuse lower gill rakers, a wide interorbital width (26.1–30.6% HL), a small eye (16.7–26.4% HL), and a short, second anal spine (25.9–37.1% HL). Two related species,J. heterolepis Bleeker from “Suriname” andJ. cantori Bleeker from Malaya, are resurrected as valid West Pacific species of Johnius (Johnius).     

Infantile hypertrophic pyloric stenosis: evaluation of three positional candidate genes, <Emphasis Type="Italic">TRPC1</Emphasis>, <Emphasis Type="Italic">TRPC5</Emphasis> and <Emphasis Type="Italic">TRPC6</Emphasis>, by association analysis and re-sequencing

Infantile hypertrophic pyloric stenosis (IHPS) is the most common inherited form of gastrointestinal obstruction in infancy with a striking male preponderance. Infants present with vomiting due to gastric outlet obstruction caused by hypertrophy of the smooth muscle of the pylorus. Two loci specific to extended pedigrees displaying autosomal dominant inheritance have been identified. A genome scan identified loci on chromosomes 11q14–q22 and Xq23–q24 which are predicted to be responsible for a subset of smaller families with IHPS demonstrating non-Mendelian inheritance. The two linked chromosomal regions both harbour functional candidate genes which are members of the canonical transient receptor potential (TRPC) family of ion channels. Both TRPC5 (Xq23–q24) and TRPC6 (11q14–q22) have a potential role in smooth muscle control and hypertrophy. Here, we report suggestive evidence for a third locus on chromosome 3q12–q25 (Z _max = 2.7, p < 0.004), a region which harbours a third TRPC gene, TRPC1. Fine mapping of all three genes using a tagSNP approach and re-sequencing identified a SNP in the promoter region of TRPC6 and a missense variant in exon 4 of TRPC6 which may be putative causal variants.     

A new species of <Emphasis Type="Italic">Rhinogobius</Emphasis> (Teleostei: Gobiidae) from the Julongjiang Basin in Fujian Province,China

Ten specimens of a new freshwater goby, Rhinogobius longyanensis were collected from the small tributary of the Julongjiang Basin, Fujian Province, southeastern China. The new species can be distinguished from all congeneric species by following unique combination of features: second dorsal fin rays I, 8; anal fin rays I, 7–8; pectoral fin rays modally 17; longitudinal scale series 30–32; predorsal scales 6–8; vertebral count 10 + 17 = 27; three parallel, deep brown oblique stripes crossing on cheek; branchiostegal membrane with 24–28 orange red spots in male; body with 5–6 deep brown blotches; pectoral fin base with a blackish brown bar. A diagnostic key to all nominal species with high vertebral counts from mainland Southeast Asia, China, Taiwan, and Japan is also provided.     

Production and characterization of intergeneric somatic hybrids between <Emphasis Type="Italic">Brassica napus</Emphasis> and <Emphasis Type="Italic">Orychophragmus violaceus</Emphasis> and their backcrossing progenies

Alien chromosome addition lines have been widely used for identifying gene linkage groups, assigning species-specific characters to a particular chromosome and comparing gene synteny between related species. In plant breeding, their utilization lies in introgressing characters of agronomic value. The present investigation reports the production of intergeneric somatic hybrids Brassica napus (2n = 38) + Orychophragmus violaceus (2n = 24) through asymmetric fusions of mesophyll protoplasts and subsequent development of B. napus-O. violaceous chromosome addition lines. Somatic hybrids showed variations in morphology and fertility and were mixoploids (2n = 51–67) with a range of 19–28 O. violaceus chromosomes identified by genomic in situ hybridization (GISH). After pollinated with B. napus parent and following embryo rescue, 20 BC₁ plants were obtained from one hybrid. These exhibited typical serrated leaves of O. violaceus or B. napus-type leaves. All BC₁ plants were partially male fertile but female sterile because of abnormal ovules. These were mixoploids (2n = 41–54) with 9–16 chromosomes from O. violaceus. BC₂ plants showed segregations for female fertility, leaf shape and still some chromosome variation (2n = 39–43) with 2–5 O. violaceus chromosomes, but mainly containing the whole complement from B. napus. Among the selfed progenies of BC₂ plants, monosomic addition lines (2n = 39, AACC + 1O) with or without the serrated leaves of O. violaceus or female sterility were established. The complete set of additions is expected from this investigation. In addition, O. violaceus plants at diploid and tetraploid levels with some variations in morphology and chromosome numbers were regenerated from the pretreated protoplasts by iodoacetate and UV-irradiation. Z. Zhao and T. Hu make equal contributions to this work.     

A new species of the genus <Emphasis Type="Italic">Triplophysa</Emphasis> (Nemacheilinae: Balitoridae), <Emphasis Type="Italic">Triplophysa jianchuanensis</Emphasis> sp. nov,from Yunnan,China

A new loach species, named Triplophysa jianchuanensis, has been recognized in collections from Jianchuan in Dali Prefecture, Yunnan, China. Triplophysa jianchuanensis can be distinguished from the other Triplophysa species by the following combination of characteristics: smooth skin, scaleless; lateral line complete; head long (26.7–29.8% of SL); eyes large (25.9–31.0% of dorsal head length); snout shorter than postorbital length (26.9–30.1% of HL); middle of lower lip interrupted and forming a pair of furrows; lower jaw spoon-like with obtuse edge; posterior chamber of air bladder completely degenerated; intestines short, bending in zigzag-shape behind stomach; pelvic-fin tip not reaching anus; caudal fin emarginate; branched rays of dorsal fin 7; branched rays of caudal fin 16. A key to the known species of Triplophysa from the Lancangjiang River is provided.     

Benzyladenine Promotes Flowering in <Emphasis Type="Italic">Doritaenopsis</Emphasis> and <Emphasis Type="Italic">Phalaenopsis</Emphasis> Orchids

Two experiments were performed to determine how application of the cytokinin benzyladenine (BA) influenced flowering in Doritaenopsis and Phalaenopsis orchid clones. In the first experiment, two vegetative orchid clones growing in 15-cm pots were transferred from a 28°C greenhouse that inhibited flowering to a 23°C greenhouse for flower induction (day 0). A foliar spray (0.2 L m⁻²) containing BA at 100, 200, or 400 mg L⁻¹ or 25, 50, or 100 mg L⁻¹ each of BA and gibberellins A₄ + A₇ (BA+GA) was applied on days 0, 7, and 14. Plants treated with BA alone at 200 or 400 mg L⁻¹ had a visible inflorescence 3–9 days earlier and had a mean of 0.7–3.5 more inflorescences and 3–8 more flowers per plant than nontreated plants. The application of BA+GA had no effect on inflorescence number and total flower number at the rates tested. In the second experiment, three orchid clones received a single foliar spray of BA at 200 mg L⁻¹ at six time points relative to time of transfer from 29°C to 23°C (−1, 0, +1, +2, +4, or +6 weeks). A separate group of plants received a BA application at week 0 but was maintained at 29°C. Inflorescence number was greatest in all three orchid clones when plants were treated with BA 1 week after the temperature transfer. Plants that were sprayed with BA and maintained at 29°C did not initiate inflorescences. The promotion of flowering by the application of BA suggests that cytokinins at least partially regulate inflorescence initiation of Doritaenopsis and Phalaenopsis, but its promotion is conditional and BA application cannot completely substitute for an inductive low temperature.     

A new species of deepwater cardinalfish, <Emphasis Type="Italic">Epigonus mayeri</Emphasis>, from the eastern Central Atlantic,and redescription of <Emphasis Type="Italic">Epigonus heracleus</Emphasis> Parin and Abramov 1986 (Perciformes: Epigonidae)

A new epigonid fish, Epigonus mayeri, is described on the basis of two specimens (109.7–113.8 mm in standard length: SL) from off Angola, and Epigonus heracleus Parin and Abramov 1986 is redescribed on the basis of 12 additional specimens with type specimens from off eastern New Zealand. These species belong to a subgroup of Epigonus, known as the “Epigonus robustus group,” which have a pungent opercular spine and VII-I, 9 dorsal-fin rays. The new species differs from other species of the group by having a sharp-pointed mustache-like process, presence of a rib on the last abdominal vertebra, vertebrae 10 + 15, tongue toothless, pyloric caeca 5, pectoral fin reaching to vertical line from anus (length 22.2–23.6% SL), orbital diameter 16.4–17.0% SL, head length 37.8–38.0% SL, and lower jaw length 16.7–17.0% SL.     

Edible oil degradation by using yeast coculture of <Emphasis Type="Italic">Rhodotorula pacifica</Emphasis> ST3411 and <Emphasis Type="Italic">Cryptococcus laurentii</Emphasis> ST3412

To develop a microbial treatment of edible oil-contaminated wastewater, microorganisms capable of rapidly degrading edible oil were screened. The screening study yielded a yeast coculture comprising Rhodotorula pacifica strain ST3411 and Cryptococcus laurentii strain ST3412. The coculture was able to degrade efficiently even at low contents of nitrogen ([NH₄–N] = 240 mg/L) and phosphorus sources ([PO₄–P] = 90 mg/L). The 24-h degradation rate of 3,000 ppm mixed oils (salad oil/lard/beef tallow, 1:1 w/w) at 20°C was 39.8% ± 9.9% (means ± standard deviations of eight replicates). The highest degradation rate was observed at 20°C and pH 8. In a scaled-up experiment, the salad oil was rapidly degraded by the coculture from 671 ± 52.0 to 143 ± 96.7 ppm in 24 h, and the degradation rate was 79.4% ± 13.8% (means ± standard deviations of three replicates). In addition, a repetitive degradation was observed with the cell growth by only pH adjustment without addition of the cells.     

<Emphasis Type="Italic">Salinarchaeum laminariae</Emphasis> gen. nov., sp. nov.: a new member of the family <Emphasis Type="Italic">Halobacteriaceae</Emphasis> isolated from salted brown alga <Emphasis Type="Italic">Laminaria</Emphasis>

Halophilic archaeal strains R26^T and R22 were isolated from the brown alga Laminaria produced at Dalian, Liaoning Province, China. Cells from the two strains were pleomorphic rods and Gram negative, and colonies were red pigmented. Strains R26^T and R22 were able to grow at 20–50°C (optimum 37°C) in 1.4–5.1 M NaCl (optimum 3.1–4.3 M) at pH 5.5–9.5 (optimum pH 8.0–8.5) and neither strain required Mg²⁺ for growth. Cells lyse in distilled water and the minimum NaCl concentration required to prevent cell lysis was 8% (w/v) for strain R26^T and 12% (w/v) for strain R22. The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and minor phosphatidylglycerol sulfate; glycolipids were not detected. Phylogenetic analyses based on 16S rRNA genes and rpoB′ genes revealed that strains R26^T and R22 formed a distinct clade with the closest relative, Natronoarchaeum mannanilyticum. The DNA G+C content of strains R26^T and R22 was 65.8 and 66.4 mol%, respectively. The DNA–DNA hybridization value between strains R26^T and R22 was 89%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that the strains R26^T and R22 represent a novel species in a new genus within the family Halobacteriaceae, for which the name Salinarchaeum laminariae gen. nov., sp. nov. is proposed. The type strain is R26^T (type strain R26^T = CGMCC 1.10590^T = JCM 17267^T, reference strain R22 = CGMCC 1.10589).     

A new species of the genus <Emphasis Type="Italic">Careproctus</Emphasis> (Liparidae) from the Sea of Japan

A new snailfish, Careproctus notosaikaiensis, is described on the basis of 6 specimens collected from the central part of the Sea of Japan, off Saikai, western coast of the Noto Peninsula, Ishikawa, Japan. The new species is distinguished from other congeners by the following combination of the characters: vertebrae 57–58, 10–12 abdominal and 46–48 caudal; dorsal-fin rays 52; anal-fin rays 46–47; principal caudal-fin rays 10; pectoral-fin rays 35–37; pectoral fin with a deep notch; proximal pectoral radials 4 (3 + 1), round; gill slit extending to pectoral-fin ray 4–7; teeth strongly trilobed; pleural ribs 2 pairs; 2 suprabranchial pores; chin pores paired in the same pit; pyloric caeca 20–29; dorsal and anal fins with dark margins and stomach black in preserved specimens.     

Valsartan Orodispersible Tablets: Formulation, <Emphasis Type="Italic">In vitro</Emphasis>/<Emphasis Type="Italic">In vivo</Emphasis> Characterization

Valsartan orodispersible tablets have been developed at 40-mg dose, with the intention of facilitating administration to patients experiencing problems with swallowing and hopefully, improving its poor oral bioavailability. Work started with selecting drug compatible excipients depending on differential scanning calorimetric analysis. A 3³ full factorial design was adopted for the optimization of the tablets prepared by freeze-drying technique. The effects of the filler type, the binder type, and the binder concentration were studied. The different tablet formulas were characterized for their physical properties, weight variation, disintegration time, surface properties, wetting properties, and in vitro dissolution. Amongst the prepared 27 tablet formulas, formula number 6 (consisting of 4:6 valsartan:mannitol and 2% pectin) was selected to be tested in vivo. Oral bioavailability of two 40 mg valsartan orodispersible tablets was compared to the conventional commercial tablets after administration of a single dose to four healthy volunteers. Valsartan was monitored in plasma by high-performance liquid chromatography. The apparent rate of absorption of valsartan from the prepared tablets (C _max = 2.879 μg/ml, t _max = 1.08 h) was significantly higher than that of the conventional tablets (C _max = 1.471 μg/ml, t _max = 2.17 h), P ≤ 0.05. The relative bioavailability calculated as the ratio of mean total area under the plasma concentration–time curve for the orodispersible tablets relative to the conventional ones was 135%. The results of the in vivo study revealed that valsartan orodispersible tablets would be advantageous with regards to improved patient compliance, rapid onset of action, and increase in bioavailability.     

Redescription of <Emphasis Type="Italic">Bregmaceros mcclellandi</Emphasis> Thompson, 1840 (Gadiformes: Bregmacerotidae)

 This study redescribes Bregmaceros mcclellandi Thompson, 1840, based on one specimen (74.4 mm SL) from the Bay of Bengal and 66 specimens (30.0–84.7 mm SL) from Mumbai (Bombay), India, because the type specimens have apparently been lost. The present specimens are characterized by having black dorsal, pectoral, and caudal fins and show the following morphology: caudal fin slightly forked; body chromatophores present mainly at the dorsal part; no scales on cheek; vertebrae 52–55 (13–15 + 38–41); dorsal rays 52–59; anal rays 54–60; pectoral rays 18–20; caudal rays 27–31 (principal rays 14); transverse scales 14–15. In the 66 Mumbai specimens, it was confirmed that the distinctive black fin pigmentation developed sequentially with growth, with complete pigmentation first on the anterior lobe of the dorsal fin, then simultaneously on the posterior lobe of the dorsal fin, the caudal fin, and the pectoral fin, and last, on the anal fin. This species is known only from the Arabian Sea, Bay of Bengal, and Gulf of Thailand. A review of 16 nominal Bregmaceros species indicates that, besides B. mcclellandi, the distinctive dark fin pigmentation is found in B. atripinnis (Tickell), B. atlanticus Goode and Bean, B. japonicus Tanaka, and B. lanceolatus Shen. B. atripinnis is considered a junior synonym of B. mcclellandi, and the others are clearly distinct from B. mcclellandi. Comments are made on some of the characters to more fully characterize the species and for reference in future revisionary and phylogenetic studies. Received: June 17, 2002 / Revised: December 2, 2002 / Accepted: December 24, 2002     

Purification and characterization of chitinases from <Emphasis Type="Italic">Paecilomyces</Emphasis><Emphasis Type="Italic">variotii</Emphasis> DG-3 parasitizing on <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> eggs

Two extracellular chitinases were purified from Paecilomyces variotii DG-3, a chitinase producer and a nematode egg-parasitic fungus, to homogeneity by DEAE Sephadex A-50 and Sephadex G-100 chromatography. The purified enzymes were a monomer with an apparent molecular mass of 32 kDa (Chi32) and 46 kDa (Chi46), respectively, and showed chitinase activity bands with 0.01% glycol chitin as a substrate after SDS-PAGE. The first 20 and 15 N-terminal amino acid sequences of Chi32 and Chi46 were determined to be Asp-Pro-Typ-Gln-Thr-Asn-Val-Val-Tyr-Thr-Gly-Gln-Asp-Phe-Val-Ser-Pro-Asp-Leu-Phe and Asp-Ala-X-X-Tyr-Arg-Ser-Val-Ala-Tyr-Phe-Val-Asn-Trp-Ala, respectively. Optimal temperature and pH of the Chi32 and Chi46 were found to be both 60°C, and 2.5 and 3.0, respectively. Chi32 was almost inhibited by metal ions Ag⁺ and Hg²⁺ while Chi46 by Hg²⁺ and Pb²⁺ at a 10 mM concentration but both enzymes were enhanced by 1 mM concentration of Co²⁺. On analyzing the hydrolyzates of chitin oligomers [(GlcNAc) _n , n = 2–6)], it was considered that Chi32 degraded chitin oligomers as an exo-type chitinase while Chi46 as an endo-type chitinase.     

Improved clonal propagation of <Emphasis Type="Italic">Spilanthes</Emphasis><Emphasis Type="Italic">acmella</Emphasis> Murr. for production of scopoletin

A reproducible protocol for clonal propagation of Spilanthes acmella has been established. Routinely, the cultures were established in spring (January–April) season because of the highest aseptic culture establishment and high frequency shoot proliferation. Incorporation of 5 μM N⁶-benzyladenine (BA) to Murashige and Skoog (MS) basal medium showed 100% bud-break and promoted multiple shoot proliferation in cultures. Interestingly, a higher concentration of BA (7–15 μM) promoted stunted shoots with pale leaves while a lower concentration (1–3 μM) resulted in shoots with long internodes and excessive adventitious root proliferation from all over their surface. For recurrent shoot multiplication, single node segments from in vitro-developed shoots were excised and cultured on MS + BA (5 μM) medium where 20.3-fold shoot multiplication was achieved every 5 weeks. Finally, these shoots were successfully rooted on half-strength MS medium (major salts reduced to half-strength) with 50 g l⁻¹ sucrose, with a frequency of 100%. Transplantation survival of micropropagated plants was 88.9%. Additionally, accumulation of scopoletin, a phytoalexin, was revealed for the first time in the uninfected leaves of Spilanthes. Further, the quantitative estimation by HPLC with a fluorescence detector showed that the amounts of scopoletin content (0.10 μg g⁻¹ DW) in the leaves of micropropagated plants are comparable to those of field-grown mother plants. The study thus signifies the effectiveness of in vitro methodology for true-to-type plant regeneration of Spilanthes and their later utility for biosynthesis and constant production of scopoletin throughout the year.     

Pulse perturbations from bacterial decomposition of <Emphasis Type="Italic">Chrysaora</Emphasis><Emphasis Type="Italic">quinquecirrha</Emphasis> (Scyphozoa: Pelagiidae)

Bacteria decomposed damaged and moribund Chrysaora quinquecirrha Desor, 1848 releasing a pulse of carbon and nutrients. Tissue decomposed in 5–8 days, with 14 g of wet biomass exhibiting a half-life of 3 days at 22°C, which is 3× longer than previous reports. Decomposition raised mean concentrations of organic carbon and nutrients above controls by 1–2 orders of magnitude. An increase in nitrogen (16,117 μg l⁻¹) occurred 24 h after increases in phosphorus (1,365 μg l⁻¹) and organic carbon (25 mg l⁻¹). Cocci dominated control incubations, with no significant increase in numbers. In incubations of tissue, bacilli increased exponentially after 6 h to become dominant, and cocci reproduced at a rate that was 30% slower. These results, and those from previous studies, suggested that natural assemblages may include bacteria that decompose medusae, as well as bacteria that benefit from the subsequent release of carbon and nutrients. This experiment also indicated that proteins and other nitrogenous compounds are less labile in damaged medusae than in dead or homogenized individuals. Overall, dense patches of decomposing medusae represent an important, but poorly documented, component of the trophic shunt that diverts carbon and nutrients incorporated by gelatinous zooplankton into microbial trophic webs.     

Reclassification of Two <Emphasis Type="Italic">Peronospora</Emphasis> Species Parasitic on <Emphasis Type="Italic">Draba</Emphasis> in <Emphasis Type="Italic">Hyaloperonospora</Emphasis> Based on Morphological and Molecular Phylogenetic Data

On the family Brassicaceae, the causal agent responsible for downy mildew disease was originally regarded as a single species, Peronospora parasitica (now under Hyaloperonospora), but it was recently reconsidered to consist of many distinct species. In this study, 11 specimens of Peronospora drabae and P. norvegica parasitic on the genus Draba were investigated morphologically and molecularly. Pronounced differences in conidial sizes (P. drabae: 14–20 × 12.5–15.5 μm; P. norvegica: 20–29 × 15.5–22 μm) and 7.8% sequence distance between their ITS1-5.8S-ITS2 rDNA sequences confirmed their status as distinct species. Based on ITS phylogeny and morphology (monopodially branching conidiophores, flexuous to sigmoid ultimate branchlets, hyaline conidia and lobate haustoria), the two species unequivocally belong to the genus Hyaloperonospora and not to Peronospora to which they were previously assigned. Therefore, two new combinations, Hyaloperonospora drabae and H. norvegica, are proposed. The two taxa are illustrated and compared using the type specimen for H. norvegica and authentic specimens for H. drabae, which is lectotypified.