Pethia sanjaymoluri,a new species of barb (Teleostei: Cyprinidae) from the northern Western Ghats,India

Pethia sanjaymoluri, a new cyprinid, is described from the Pavana and Nira tributaries of Bhima River, Krishna drainage, Maharashtra, India. It can be distinguished from congeners by a combination of characteristics that includes an incomplete lateral line, absence of barbels, upper lip thick and fleshy, 23–25 lateral series scales, 7–12 lateral‐line pored scales, 10 predorsal scales, 11–14 prepelvic scales, 17–20 pre‐anal scales, 4½ scales between dorsal‐fin origin and lateral line, four scales between lateral line and pelvic‐fin origin, 8–15 pairs of serrae on distal half of dorsal‐fin spine, 12–14 branched pectoral‐fin rays, 4 + 26 total vertebrae, 4 + 5 predorsal vertebrae, 4 + 13 abdominal vertebrae, 13 caudal vertebrae and a unique colour pattern comprising a humeral spot positioned below the lateral line and encompassing the third and fourth lateral‐line scales and one scale below, one caudal spot on 17th–21st lateral‐line scales with a yellow hue on its anterior side and apical half of dorsal fin studded with melanophores making the fin tip appear black. Genetic analysis based on the mitochondrial cytochrome b gene sequence suggests that the species is distinct from other known species of Pethia for which data are available.     

Microsatellite markers from a commercially important South‐east Asian cyprinid,the silver barb (Barbodes gonionotus Bleeker)

Barbodes gonionotus (Puntius gonionotus/javanicus, Bleeker) is a cyprinid fish, endemic to South‐east Asia and is of great importance in both wild fisheries and in aquaculture in the region. It is a common to dominant species in the middle and lower stretches of medium and large rivers. Five highly polymorphic microsatellite loci are described (H_O = 0.7–0.9, number of alleles per locus = 10–37), which will be of use both in population genetic studies and for the assessment of true effective breeding population sizes in aquaculture stocks.     

Sperm cryopreservation of the critically endangered olive barb (Sarpunti) Puntiussarana (Hamilton, 1822)

The present study focused on development of a sperm cryopreservation protocol for the critically endangered olive barb Puntiussarana (Hamilton, 1822) collected from two stocks within Bangladesh and reared in the Fisheries Field Laboratory, Bangladesh Agricultural University (BAU). The sperm were collected in Alsever’s solution prepared at 296 mOsmol kg⁻¹. Sperm were activated with distilled water (24 mOsmol kg⁻¹) to characterize motility. Maximum motility (90%) was observed within 15 s after activation, and sperm remained motile for 35 s. Sperm activation was evaluated in different osmolalities and motility was completely inhibited when osmolality of the extender was ?287 mOsmol kg⁻¹. To evaluate cryoprotectant toxicity, sperm were equilibrated with 5%, 10% and 15% each of dimethyl sulfoxide (DMSO) and methanol. Sperm motility was noticeably reduced within 10 min, when sperm were equilibrated with 15% DMSO, indicating acute toxicity to spermatozoa and therefore this concentration was excluded in further trials. Sperm were cryopreserved using DMSO at concentrations of 5% and 10% and methanol at 5%, 10% and 15%. The one-step freezing protocol (from 5 °C to −80 °C at 10 °C/min) was carried out in a computer-controlled freezer (FREEZE CONTROL® CL-3300; Australia) and 0.25-ml straws containing spermatozoa were stored in liquid nitrogen for 7–15 days at −196 °C. The highest motility in thawed sperm 61 ± 8% (mean ± SD) was obtained with 10% DMSO. The fertilization and hatching rates were 70% and 37% for cryopreserved sperm, and 72% and 62% for fresh sperm. The protocol reported here can be useful for hatchery-scale production of olive barb. The use of cryopreserved sperm can facilitate hatchery operations, and can provide for long-term conservation of genetic resources to contribute in the recovery of critically endangered fish such as the olive barb.     

Early embryonic development of the mayfly Ephemera japonica McLachlan (Insecta: Ephemeroptera,Ephemeridae)

In the newly laid egg of the mayfly Ephemera japonica, an egg nucleus (oocyte nucleus) at metaphase of the first maturation division is in the polar plasm at the mid-ventral side of the egg, and a male pronucleus lies in the periplasm beneath a micropyle situated just opposite the polar plasm or at the mid-dorsal side of egg. The maturation divisions are typical. An extensive and circuitous migration of the male pronucleus is involved in the fertilization process: it first moves anteriad in the periplasm from beneath the micropyle to the anterior pole of the egg and then turns posteriad in the yolk along the egg's long axis to the site of syngamy, near the center of the egg. Cleavage is superficial. The successive eight cleavages, of which the first five are synchronized, result in the formation of the blastoderm, and about ten primary yolk cells remain behind in the yolk. Even in the newly formed blastoderm, the thick embryonic posterior half and the thin extraembryonic anterior half areas are distinguished: the former cells are concentrated at the posterior pole of the egg to form the germ disc, and the latter cells become more flattened, forming serosa. Time-lapse VTR observations reveal a yolk stream that is in accord with the migration of the male pronucleus in time and direction. The yolk stream is also generated in activated unfertilized eggs, and it is probable that the migration of the male pronucleus in association with the fertilization may be directed by the yolk stream. J. Morphol. 238:327–335, 1998. © 1998 Wiley-Liss, Inc.     

Identification and characterization of microsatellite DNA markers developed in an endangered cyprinid of the Mekong River,the seven‐line barb (Probarbus jullieni Sauvage, 1880)

Microsatellite markers for an endangered cyprinid species of the Mekong River, the seven‐line barb (Probarbus jullieni Sauvage, 1880) were developed from the wild caught samples using (GT)₁₅ probe. The number of alleles per locus ranged from seven to 16. The expected heterozygosities ranged from 0.47 to 0.91. Also, these primers were successfully amplified in the two closely related species, P. labeamajor and P. labeaminor. These markers have proven to be very useful for the population genetic structure study in this species and other related cyprinids.     

Location of micropyles and early embryonic development of the two-spotted cricket Gryllus bimaculatus (Insecta, Orthoptera)

Early embryogenesis of the two-spotted cricket Gryllus bimaculatus was examined by scanning electron microscopy and several fluorescence staining methods, with special reference to these four issues: (i) the location of micropyles; (ii) the transfer of the female pronucleus following meiosis; (iii) the timing of cellularization; and (iv) the process of the germ primordium formation. Between two and four micropyles lie in the mid-ventral region of the egg. The egg nucleus is at the mid-dorsal periphery of the new laid egg, and meiosis resumes and is completed there. The female pronucleus moves to the mid-ventral side, and fertilization occurs there. Energid starts to proliferate and migrates to the periphery of the egg, initiating blastoderm formation. Actin caps surround each superficial nucleus. Cellularization occurs during the blastoderm stage. At a late blastoderm stage, nuclei aggregate in both the posterolateral patch-like regions of the egg to form a germ primordium. The germ primordium looks like a pair of dumbbells. Both the patches shift towards the ventral side and fuse into a germ primordium. The germ primordium contracts to produce a clearly delineated germ band. Observations on distribution patterns of F-actin indicate that, all through the process, the germ primordium retains that unity, and is not separated into two parts.     

Intercalibration of hydroacoustic and mark–recapture methods for assessing the spawning population size of a threatened fish species

Hydroacoustic counting and a three-year mark–recapture study with passive integrated transponders (PIT tags) were used to estimate the size of a spawning population of nase Chondrostoma nasus , a threatened potamodromous cyprinid that undertakes annual spawning migrations into a tributary of the Danube River. In 2005, the estimates of the size of the spawning population from the hydroacoustic counts ( N = 2234, 95% CL 1929–2538) and from the Jolly–Seber model ( N = 1198, 95% CL 461–5842) corresponded well. Estimates from the jackknife-estimator based on the hydroacoustic counts yielded slightly higher values ( N = 2783, 95% CL 2529–3037), but were still in the same order of magnitude as those from the hydroacoustic and mark–recapture approach. At low run-size, hydroacoustic counting was more time consuming and technically demanding than mark–recapture studies. At the same time, it was non-invasive, provided real-time data on a fine temporal scale, and estimates showed less variability than the Jolly–Seber model. Mark–recapture of fish in spawning streams involved substantial disturbance at a sensitive stage of the life cycle. Hence, hydroacoustics is highly suited for population estimates of threatened potamodromous fishes, where interference needs to be minimized.     

Morphometric study of embryonic development of Macrobrachium borellii (Arthropoda: Crustacea)

Summary

Embryo morphometry and developing time from just-laid eggs until hatching were described in the palaemonid prawn, Macrobrachium borellii, using a rapid non-invasive staging method. Embryos were kept in the laboratory under controlled conditions and development divided into seven stages according to major morphological characteristics. This lecithotrophic, freshwater shrimp has a highly abbreviated type of hatching development after 39 ± 2 days as postlarvae at 24°C. Morphometry was recorded using a stereoscopic microscope with an image analyzer. Area, perimeter, maximum and minimum diameters and shape were measured in yolk sac, egg-coat and eye, respectively, and they were statistically selected as the best to define the stages. Eggs are ovoid with a maximum diameter that varies from the moment of oviposition to the time of hatching from 1.5 to 2.0 mm, respectively. Water content and egg size increase along with development, whereas egg shape only varies just before hatching when the egg becomes strongly ovoid. Egg coat and eye variables significantly increase as the embryo develops while all yolk variables decrease as the embryo consumes the vitellus. Yolk represents more than 95% of the egg at the time of oviposition, falling to 22% by the time of hatching. The major yolk area decrease is observed between stages 4 and 5, which is coincident with a marked increase in the catabolism. Using only egg coat, yolk and eye shape and maximum diameter, a researcher can straightforwardly identify a developing stage with an accuracy ranging from 70 to 100%. This tool may be employed in other species provided they have transparent chorion.     

Reproductive biology of the endemic cyprinid fish Hypselobarbus thomassi (Day, 1874) from Kallada River in the Western Ghats,India

The study provides new information on the reproductive biology of Hypselobarbus thomassi (Day, 1874) an endemic cyprinid fish in the Western Ghats–a biodiversity hotspot of India. A total of 384 specimens (males = 244 and females = 140) were collected monthly from April 2009 to March 2011 from Kallada River of Southern part of Western Ghats, India. Monthly sex ratio indicated that males predominated in the fishery, and an overall sex ratio was significantly different from the expected value of 1:1 (male:female = 1:0.57, x² = 28.17, p < .01). Based on the monthly analysis of gonadosomatic index (GSI) and the proportion of mature individuals, the spawning season of H. thomassi occurred between May to October with a peak in June to August. The length at first maturity (L₅₀) recorded was 290 mm TL in males and 330 mm TL in females. The absolute fecundity ranged between 305 (234 mm TL) to 1,089 (414 mm TL) and relative fecundity oscillated between 77.3–220.69 oocytes/g of fish. The fecundity established a linear relationship with total length, body weight and ovary weight. The findings of this study would be very effective to impose sustainable conservation plan for this threatened species in Kallada River and other Western Ghats river systems.     

解磷菌株B25的筛选、鉴定及其解磷能力

从安徽省铜陵市铜官山尾矿库木贼根际分离筛选出多株解磷细菌,经过多次筛选纯化获得一株解磷能力较好的菌株B25.采用透射电镜观察和DNA分子技术,确定此菌株属于芽孢杆菌属.研究了解磷菌株B25在培养168 h内的解磷能力、溶液pH值以及菌株生长量的变化情况,并比较了B25在不同条件下的解磷能力.结果表明：解磷菌株B25的解磷能力与溶液pH值之间存在微弱的相关性,在碳源为葡萄糖、初始pH值为7.0、培养温度为30 ℃时解磷效果较好.     

Distribution of cells containing FMRFamide-related molecules in the embryonic development of Ophryotrocha labronica (Polychaeta: Dorvilleidae)

Abstract. The timing and spatial distribution of cells containing FMRFamide-related molecules in the embryogenesis of the polychaete Ophryotrocha labronica were studied immunocytochemically. FMRFamide-like molecules emerge early during embryonic development. They are found at the one-cell stage, are asymmetrically distributed in the first phases of cleavage, associated with gastrular movements, and label the central nervous system morphogenesis. Moreover, during embryogenesis, the pattern of gut cells with the FMRFamide-like phenotype that is present in adults is already established. The early occurrence of FMRFamide-like molecules in O. labronica suggests that these molecules are involved as pre-nervous growth signals in the regulation of basic neuronal cell behaviors.     

Induced spawning and embryonic and early larval development of bonefish (Albula vulpes)

Bonefish (Albula vulpes L.) are a highly prized sport fish. Despite their economic importance, populations in the Florida Keys and Caribbean are in decline, with the early life history undescribed. Injections of carp pituitary extract into A. vulpes during the advanced stages of ovarian development induced ovulation and spawning. Embryos were sampled hourly until hatching into undeveloped, yolk-sac leptocephalus larvae. These larvae survived 56 h post-hatch, when myomeres and eyes were developing but not the mouth. These results inform future research on the reproduction and early life history of A. vulpes.     

Effect of a juvenoid on embryonic development of Earias fabia (Stoll) (Lepidoptera: Noctuidae)

Eggs of Earias fabia were treated with three different quantities of a juvenoid, 6–7 epoxy-3-ethyl-1-(p-ethyl phenoxy)-7 methylnonane at 5 distinct stages. These were: (a) before blastoderm formation; (b) before gastrulation; (c) before segmentation; (d) before blastokinesis; and (e) just after blastokinesis. The effect on embryonic development was studied. If sufficient hormone is administered early enough, development may be arrested at stages as early as blastoderm formation. With moderate and smaller doses, developmental arrest mostly occurred at certain well-defined stages, such as germ-band formation; before blastokinesis; post blastokinesis; when the embryo assumes its larval form; at the time of chitin formation and during embryonic moulting. Observations also bring in focus the importance of the quantity of hormone and its relationship to the speed of its action. Toxic action is apparent when eggs are treated with relatively larger quantities of the compound: application of juvenoid to younger stages causes fewer deaths.     

Early development of milkfish: effects of salinity on embryonic and larval metabolism, yolk absorption and growth

During embryogenesis of Chanos chanos , more than half of the yolk was consumed and the majority of it was converted into larval tissue. Salinity affected both yolk absorption and embryonic and larval growth. Larvae hatched in 20% had larger yolk reserves but were smaller and grew more slowly than larvae in 35 and 50%. Larvae hatched in 35 and 50% had equal amounts of yolk but those from 35% were larger. Oxygen consumption rates increased during development (from 0.06 ± 0.01 μl O₂ egg^–1 h^–1 by blastulae to 0.37 ± 0-01 μl O₂ egg^–1 h^–1 by prehatch embryos and 0–43 ± 0–03 μl O₂ larva ^–1 h ^–1 by newly-hatched larvae) and were significantly affected by salinity. Eggs and yolk-sac larvae incubated in 35% consumed more oxygen than those in the low and high salinities. Salinity affected both the rate and pattern of yolk utilization but salinity-related differences in metabolism, yolk absorption, and growth were not related directly to the osmotic gradient. Low salinity retarded yolk absorption while high salinity reduced yolk utilization efficiencies. Differences in oxygen consumption rates were probably related to variations in the relative amounts of metabolically active embryonic and larval tissue and/or higher activity levels rather than differential osmoregulatory costs. 35% is probably the most suitable salinity for incubation and larval rearing of milkfish.     

Early embryonic development of the freshwater shrimp Caridina multidentata (Crustacea,Decapoda, Atyidae)

Decapod crustaceans show a great diversity of developmental modes at all levels. In particular, early cleavage varies from total via mixed to superficial modes and from determinate cleavage with a stereotyped pattern to indeterminate cleavage. However, the ground pattern of early decapod development is not clear. To address this problem, we studied the early embryonic development of the caridean shrimp Caridina multidentata with a combination of confocal laser scanning microscopy, scanning electron microscopy, 4D microscopy and 3-D reconstruction software. Despite a yolk-rich egg, the cleavage is holoblastic and shows a distinct pattern of blastomere arrangement, characterized by two interlocking cell bands. This resembles the conditions in dendrobranchiate shrimps, which most likely are the sister group to Pleocyemata to which C. multidentata belongs. Hence, our results offer the possibility to assume total cleavage with blastomeres arranged in two interlocking cell bands as ancestral cleavage mode for Decapoda.     

A New Species of the Genus Tor from Yunnan, China (Teleostei: Cyprinidae)

A new species of the subfamily Barbinae, Tor yingjiangensis, is recognized from Yunnan province, China. It can be distinguished from other Tor species by the following combination of characters: last simple dorsal-fin ray osseous and non-serrated; no forward directed predorsal procumbent spine; head length considerably longer than body depth; no tubercles on the snout or sides of the face; 18–20 gill rakers on the outside of first gill arch; 24–26 lateral line scales; median lobe of lower lip short, its posterior margin truncate, not extending to the vertical across the inner corners of the mouth; and the condition of the lower lip consistent in individuals of different sizes. The new species has been misidentified previously as Tor putitora (Hamilton, F. 1822. An account of the fishes found in the River Ganges and its Branches. Edinburgh & London. 405 pp), which occurs in the Ganges and Indus River basins. The new species can easily be distinguished from T. putitora by having 3–3.5 (vs. 2.5) scales from lateral line to pelvic-fin origin, shorter caudal peduncle length (13.0% vs. 17.2% of standard length), lesser body depth (26.4% vs. 24.0% of standard length) and longer caudal peduncle depth (12.0% vs. 10.9% of standard length), no longitudinal stripe present along side of body, and eyes visible in ventral view of head.     

Early embryonic development of the head region of Gryllus assimilis Fabricius, 1775 (Orthoptera,Insecta)

We report our investigations on the embryonic development of Gryllus assimilis, with particular attention to the head. Significant findings revealed with scanning electron microscopy (SEM) images include: (1) the pre-antennal lobes represent the anterior-most segment that does not bear any appendages; (2) each of the lobes consists of central and marginal regions; (3) the central region thereof develops into the protocerebrum and the optic lobes, whereas the marginal region thereof becomes the anterior portion of the head capsule; (4) the initial position of the antennal segment is posterior to the mouth region; (5) appendage anlagen are transitorily present in the intercalary segment, and they later vanish together with the segment itself; (6) a bulged sternum appears to develop from the ventral surface of the mandibular, maxillary and labial segments. Embryonic features are then compared across the Insecta and further extended to the embryos of a spider (Araneae, Chelicerata). Striking similarities shared by the anterior-most region of the insect and spider embryos lead the authors to conclude that such comparison should be further undertaken to cover the entire Euarthropoda. This will help us to understand the embryology and evolution of the arthropod head.     

Stages of embryonic development in the amphipod crustacean, Parhyale hawaiensis

Studying the relationship between development and evolution and its role in the generation of biological diversity has been reinvigorated by new techniques in genetics and molecular biology. However, exploiting these techniques to examine the evolution of development requires that a great deal of detail be known regarding the embryonic development of multiple species studied in a phylogenetic context. Crustaceans are an enormously successful group of arthropods and extant species demonstrate a wide diversity of morphologies and life histories. One of the most speciose orders within the Crustacea is the Amphipoda. The embryonic development of a new crustacean model system, the amphipod Parhyale hawaiensis, is described in a series of discrete stages easily identified by examination of living animals and the use of commonly available molecular markers on fixed specimens. Complete embryogenesis occurs in 250 h at 26 degrees C and has been divided into 30 stages. This staging data will facilitate comparative analyses of embryonic development among crustaceans in particular, as well as between different arthropod groups. In addition, several aspects of Parhyale embryonic development make this species particularly suitable for a broad range of experimental manipulations.